Tetrahydroquinoline derivatives useful as bromodomain inhibitors

ABSTRACT

Tetrahydroquinoline compounds, pharmaceutical compositions containing such compounds and their use in therapy.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is filed pursuant to 35 USC 371 as a United StatesNational Phase Application of International Patent Application SerialNo. PCT/EP2012/057113 filed on Apr. 19, 2012, which claims priority from1106750.1 filed on Apr. 21, 2011 in the United Kingdom.

FIELD OF THE INVENTION

The present invention relates to tetrahydroquinoline derivatives,pharmaceutical compositions containing such compounds and to their usein therapy.

BACKGROUND OF THE INVENTION

The genomes of eukaryotic organisms are highly organised within thenucleus of the cell. The long strands of duplex DNA are wrapped aroundan octomer of histone proteins (most usually comprising two copies ofhistones H2A, H2B H3 and H4) to form a nucleosome. This basic unit isthen further compressed by the aggregation and folding of nucleosomes toform a highly condensed chromatin structure. A range of different statesof condensation are possible, and the tightness of this structure variesduring the cell cycle, being most compact during the process of celldivision. Chromatin structure plays a critical role in regulating genetranscription, which cannot occur efficiently from highly condensedchromatin. The chromatin structure is controlled by a series of posttranslational modifications to histone proteins, notably histones H3 andH4, and most commonly within the histone tails which extend beyond thecore nucleosome structure. These modifications include acetylation,methylation, phosphorylation, ubiquitinylation, SUMOylation. Theseepigenetic marks are written and erased by specific enzymes, which placethe tags on specific residues within the histone tail, thereby formingan epigenetic code, which is then interpreted by the cell to allow genespecific regulation of chromatin structure and thereby transcription.

Histone acetylation is most usually associated with the activation ofgene transcription, as the modification loosens the interaction of theDNA and the histone octomer by changing the electrostatics. In additionto this physical change, specific proteins bind to acetylated lysineresidues within histones to read the epigenetic code. Bromodomains aresmall (˜110 amino acid) distinct domains within proteins that bind toacetylated lysine resides commonly but not exclusively in the context ofhistones. There is a family of around 50 proteins known to containbromodomains, and they have a range of functions within the cell.

The BET family of bromodomain containing proteins comprises 4 proteins(BRD2, BRD3, BRD4 and BRD-t) which contain tandem bromodomains capableof binding to two acetylated lysine residues in close proximity,increasing the specificity of the interaction. BRD2 and BRD3 arereported to associate with histones along actively transcribed genes andmay be involved in facilitating transcriptional elongation (Leroy et al,Mol. Cell. 2008 30(1):51-60), while BRD4 appears to be involved in therecruitment of the pTEF-3 complex to inducible genes, resulting inphosphorylation of RNA polymerase and increased transcriptional output(Hargreaves et al, Cell, 2009 138(1): 129-145). It has also beenreported that BRD4 or BRD3 may fuse with NUT (nuclear protein in testis)forming novel fusion oncogenes, BRD4-NUT or BRD3-NUT, in a highlymalignant form of epithelial neoplasia (French et al. Cancer Research,2003, 63, 304-307 and French et al. Journal of Clinical Oncology, 2004,22 (20), 4135-4139). Data suggests that BRD-NUT fusion proteinscontribute to carcinogensesis (Oncogene, 2008, 27, 2237-2242). BRD-t isuniquely expressed in the testes and ovary. All family members have beenreported to have some function in controlling or executing aspects ofthe cell cycle, and have been shown to remain in complex withchromosomes during cell division—suggesting a role in the maintenance ofepigenetic memory. In addition some viruses make use of these proteinsto tether their genomes to the host cell chromatin, as part of theprocess of viral replication (You et al Cell, 2004 117(3):349-60).

Japanese patent application JP2008-156311 discloses a benzimidazolederivative which is said to be a BRD2 bromodomain binding agent whichhas utility with respect to virus infection/proliferation.

Patent application WO2009084693 discloses a series ofthienotriazolodiazepiene derivatives that are said to inhibit thebinding between an acetylated histone and a bromodomain containingprotein which are said to be useful as anti-cancer agents.

PCT patent applications PCT/EP2010/06693 and PCT/EP2010/066701 disclosea series of tetrahydroquinoline derivatives that inhibit the binding ofBET family bromodomains with acetylated lysine residues.

A novel class of compounds have been found which inhibit the binding ofbromodomains with its cognate acetylated proteins, more particularly aclass of compounds that inhibit the binding of BET family bromodomainsto acetylated lysine residues. Such compounds will hereafter be referredto as “bromodomain inhibitors”.

SUMMARY OF THE INVENTION

In a first aspect of the present invention, there is provided a compoundof formula (I) or a salt thereof, more particularly a pharmaceuticallyacceptable salt thereof

In a second aspect of the present invention, there is provided apharmaceutical composition comprising a compound of formula (I) or apharmaceutically acceptable salt thereof and one or morepharmaceutically acceptable carriers, diluents or excipients.

In a third aspect of the present invention, there is provided a compoundof formula (I), or a pharmaceutically acceptable salt thereof for use intherapy, in particular in the treatment of diseases or conditions forwhich a bromodomain inhibitor is indicated.

In a fourth aspect of the present invention, there is provided a methodof treating diseases or conditions for which a bromodomain inhibitor isindicated in a subject in need thereof which comprises administering atherapeutically effective amount of a compound of formula (I) or apharmaceutically acceptable salt thereof.

In a fifth aspect of the present invention, there is provided the use ofa compound of formula (I), or a pharmaceutically acceptable salt thereofin the manufacture of a medicament for the treatment of diseases orconditions for which a bromodomain inhibitor is indicated.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a compound of formula (I) or a saltthereof.

in which

X and Y are independently CH or N provided that at least one of X and Ymust be CH;

R¹ is a group C(O)OR⁴ in which R⁴ is C₁₋₃alkyl or C₃₋₇cycloalkyl; or

R¹ is a group selected from phenyl, pyridyl, pyrazinyl and pyrimidinylsaid groups being optionally substituted by one or two substituentsselected from halogen, C₁₋₄alkyl and CN;

R² is C₁₋₄alkyl;

R³ is C₁₋₄alkyl;

R⁵ is hydrogen and R⁶ is C₁₋₄alkyl substituted by one or more hydroxy ora —NR⁷R⁸ group in which R⁷ and R⁸ are independently hydrogen or aC₁₋₄alkyl group; or

R⁵ and R⁶ together with the N to which they are attached form a 4, 5 or6 membered heterocyclyl ring optionally containing a further heteroatomselected from N, O and S, said heterocyclyl ring being optionallysubstituted by one or more C₁₋₄alkyl, hydroxyl or amino groups; and

m is 0, 1 or 2.

In one embodiment the invention provides compounds of formula (I) withcis relative stereochemistry across the tetrahydroquinoline ring inrespect of the substituents in the 2 and 4 position on the ring. In oneembodiment the compound of formula (I) or a salt thereof is the (2S,4R)enantiomer.

In one embodiment X and Y are both CH. In a further embodiment X is CHand Y is N. In a further embodiment X is N and Y is CH.

In one embodiment R¹ is a group C(O)OR⁴ in which R⁴ is isopropyl.

In one embodiment R¹ is selected from

In one embodiment R² is methyl.

In one embodiment R³ is methyl.

In one embodiment m is 0. In a further embodiment m is 1. In a furtherembodiment m is 2.

In one embodiment R⁵ is hydrogen and R⁶ is selected from

In one embodiment R⁵ and R⁶ together with the N to which they areattached form a 4, 5 or 6 membered heterocyclyl ring selected fromazetidinyl, pyrrodinyl, piperazinyl, piperidinyl and morpholinyl saidheterocyclyl ring being optionally substituted by one or more (e.g twoor three) C₁₋₄alkyl (such as methyl), hydroxyl, or amino groups.

In a further embodiment R⁵ and R⁶ together with the N to which they areattached form a group selected from

While the embodiments for each variable have generally been listed aboveseparately for each variable, this invention is intended to include allcombinations of embodiments described hereinabove including saltsthereof.

Compounds according to the invention include Examples 1-68 as describedherein. Particular compounds of the invention include:

-   2-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)acetamide;-   6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide;    and-   6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)nicotinamide    or a salt thereof.

Throughout the present specification, unless otherwise stated:

-   -   the term “halogen” is used to describe a group selected from        fluorine, chlorine or bromine;    -   the terms “C₁₋₃alkyl”, “C₁₋₄alkyl” and “C₁₋₆alkyl” are used to        describe a group or a part of the group comprising a linear or        branched alkyl group containing from 1 to 3, 1 to 4 or 1 to 6        carbon atoms respectively. Suitable examples of such groups        include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,        t-butyl, pentyl and hexyl;    -   the term “C₃₋₇cycloalkyl” is used to describe a non-aromatic        carbocyclic ring containing at least three and at most seven        carbon atoms. Examples of C₃₋₇cycloalkyl include cyclopropyl,        cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

It will be appreciated that the present invention covers compounds offormula (I) as the free base and as salts thereof, for example as apharmaceutically acceptable salt thereof.

In one embodiment the invention relates to compounds of formula (I) or apharmaceutically acceptable salt thereof.

Because of their potential use in medicine, salts of the compounds offormula (I) are desirably pharmaceutically acceptable. Suitablepharmaceutically acceptable salts can include acid or base additionsalts. As used herein, the term ‘pharmaceutically acceptable salt’ meansany pharmaceutically acceptable salt or solvate of a compound of theinvention, which upon administration to the recipient is capable ofproviding (directly or indirectly). For a review on suitable salts seeBerge et al., J. Pharm. Sci., 66:1-19, (1977). Typically, apharmaceutically acceptable salt may be readily prepared by using adesired acid or base as appropriate. The resultant salt may precipitatefrom solution and be collected by filtration or may be recovered byevaporation of the solvent.

A pharmaceutically acceptable acid addition salt can be formed byreaction of a compound of formula (I) with a suitable inorganic ororganic acid (such as hydrobromic, hydrochloric, sulphuric, nitric,phosphoric, succinc, maleic, acetic, propionic, fumaric, citric,tartaric, lactic, benzoic, salicylic, glutamaic, aspartic,p-toluenesulfonic, benzenesulfonic, methanesulfonic, ethanesulfonic,naphthalenesulfonic such as 2-naphthalenesulfonic, or hexanoic acid),optionally in a suitable solvent such as an organic solvent, to give thesalt which is usually isolated for example by crystallisation andfiltration. A pharmaceutically acceptable acid addition salt of acompound of formula (I) can comprise or be for example a hydrobromide,hydrochloride, sulfate, nitrate, phosphate, succinate, maleate, acetate,propionate, fumarate, citrate, tartrate, lactate, benzoate, salicylate,glutamate, aspartate, p-toluenesulfonate, benzenesulfonate,methanesulfonate, ethanesulfonate, naphthalenesulfonate (e.g.2-naphthalenesulfonate) or hexanoate salt.

Other non-pharmaceutically acceptable salts, e.g. formates, oxalates ortrifluoroacetates, may be used, for example in the isolation of thecompounds of formula (I), and are included within the scope of thisinvention.

The invention includes within its scope all possible stoichiometric andnon-stoichiometric forms of the salts of the compounds of formula (I).

It will be appreciated that many organic compounds can form complexeswith solvents in which they are reacted or from which they areprecipitated or crystallized. These complexes are known as “solvates”.For example, a complex with water is known as a “hydrate”. Solvents withhigh boiling points and/or capable of forming hydrogen bonds such aswater, xylene, N-methylpyrrolidinone, methanol and ethanol may be usedto form solvates. Methods for identification of solvates include, butare not limited to, NMR and microanalysis. Solvates of the compounds offormula (I) are within the scope of the invention.

The invention includes within its scope all possible stoichiometric andnon-stoichiometric forms of the solvates of the compounds of formula(I).

The invention encompasses all prodrugs, of the compound of formula (I)or a pharmaceutically acceptable salt thereof, which upon administrationto the recipient is capable of providing (directly or indirectly) thecompound of the invention, or an active metabolite or residue thereof.Such derivatives are recognizable to those skilled in the art, withoutundue experimentation. Nevertheless, reference is made to the teachingof Burger's Medicinal Chemistry and Drug Discovery, 5^(th) Edition, Vol1: Principles and Practice, which is incorporated herein by reference tothe extent of teaching such derivatives.

The compounds of formula (I) may be in crystalline or amorphous form.Furthermore, some of the crystalline forms of the compounds of formula(I) may exist as polymorphs, which are included within the scope of thepresent invention. Polymorphic forms of compounds of formula (I) may becharacterized and differentiated using a number of conventionalanalytical techniques, including, but not limited to, X-ray powderdiffraction (XRPD) patterns, infrared (IR) spectra, Raman spectra,differential scanning calorimetry (DSC), thermogravimetric analysis(TGA) and solid state nuclear magnetic resonance (SSNMR).

Compounds described herein contain chiral atoms so that optical isomers,e.g. enantiomers or diastereoisomers may be formed. Accordingly, thepresent invention encompasses all isomers of the compounds of formula(I) whether as individual isomers isolated such as to be substantiallyfree of the other isomer (i.e. pure) or as mixtures (i.e. racemates andracemic mixtures). An individual isomer isolated such as to besubstantially free of the other isomer (i.e. pure) may be isolated suchthat less than 10%, particularly less than about 1%, for example lessthan about 0.1% of the other isomer is present.

Separation of isomers may be achieved by conventional techniques knownto those skilled in the art, e.g. by fractional crystallisation,chromatography or HPLC.

Certain compounds of formula (I) may exist in one of several tautomericforms. It will be understood that the present invention encompasses alltautomers of the compounds of formula (I) whether as individualtautomers or as mixtures thereof.

It will be appreciated from the foregoing that included within the scopeof the invention are solvates, isomers and polymorphic forms of thecompounds of formula (I) and salts thereof.

The compounds of formula (I) and salts thereof may be made by a varietyof methods, including standard chemistry. Any previously definedvariable will continue to have the previously defined meaning unlessotherwise indicated. Illustrative general synthetic methods are set outbelow and then specific compounds of formula (I) or salts thereof areprepared in the working Examples.

The present invention further provides a process for the preparation ofa compound of formula (I) or a salt thereof which comprises reacting acompound of formula (II)

in which R¹, R², R³, X, Y and m are as defined in formula (I) with acompound of formula (III) or a protected derivative thereof.NR⁵R⁶  (III)in which R⁵ and R⁶ are as defined in formula (I).

The reaction of the compounds of formula (I) and formula (II) may becarried out in a suitable organic solvent (e.g. DMF or acetonitrile)with an amine (e.g. DIPEA or triethylamine) in the presence of asuitable activating agent (e.g. HATU or TBTU).

Compounds of formula (II) can be prepared by methods described herein orby analogous methods. Compounds of formula (III) are either commerciallyavailable or can be prepared by methods described herein or by analogousprocedures thereto.

It will be appreciated by those skilled in the art that it may beadvantageous to protect one or more functional groups of the compoundsdescribed. Examples of protecting groups and the means for their removalcan be found in T. W. Greene ‘Protective Groups in Organic Synthesis’(4th edition, J. Wiley and Sons, 2006). Suitable amine protecting groupsinclude acyl (e.g. acetyl, carbamate (e.g.2′,2′,2′-trichloroethoxycarbonyl, benzyloxycarbonyl or t-butoxycarbonyl)and arylalkyl (e.g. benzyl), which may be removed by hydrolysis (e.g.using an acid such as hydrochloric acid in dioxane or trifluoroaceticacid in dichloromethane) or reductively (e.g. hydrogenolysis of a benzylor benzyloxycarbonyl group or reductive removal of a2′,2′,2′-trichloroethoxycarbonyl group using zinc in acetic acid) asappropriate. Other suitable amine protecting groups includetrifluoroacetyl (—COCF₃) which may be removed by base catalysedhydrolysis.

It will be appreciated that in any of the routes described herein, theprecise order of the synthetic steps by which the various groups andmoieties are introduced into the molecule may be varied. It will bewithin the skill of the practitioner in the art to ensure that groups ormoieties introduced at one stage of the process will not be affected bysubsequent transformations and reactions, and to select the order ofsynthetic steps accordingly.

Certain intermediate compounds of formula (II) are believed to be noveland therefore form yet a further aspect of the invention.

The compounds of formula (I) and salts thereof are bromodomaininhibitors, and thus are believed to have potential utility in thetreatment of diseases or conditions for which a bromodomain inhibitor isindicated.

The present invention thus provides a compound of formula (I) or apharmaceutically acceptable salt thereof for use in therapy. Thecompound of formula (I) or pharmaceutically salt thereof can be used inthe treatment of diseases or conditions for which a bromodomaininhibitor is indicated.

The present invention thus provides a compound of formula (I) or apharmaceutically acceptable salt thereof for use in the treatment of anydiseases or conditions for which a bromodomain inhibitor is indicated.

Also provided is the use of a compound of formula (I) or apharmaceutically acceptable salt thereof in the manufacture of amedicament for the treatment of diseases or conditions for which abromodomain inhibitor is indicated.

Also provided is a method of treating diseases or conditions for which abromodomain inhibitor is indicated in a subject in need thereof whichcomprises administering a therapeutically effective amount of compoundof formula (I) or a pharmaceutically acceptable salt thereof.

Suitably the subject in need thereof is a mammal, particularly a human.

As used herein, the term “effective amount” means that amount of a drugor pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal or human that is being sought, forinstance, by a researcher or clinician. Furthermore, the term“therapeutically effective amount” means any amount which, as comparedto a corresponding subject who has not received such amount, results inimproved treatment, healing, prevention, or amelioration of a disease,disorder, or side effect, or a decrease in the rate of advancement of adisease or disorder. The term also includes within its scope amountseffective to enhance normal physiological function.

Bromodomain inhibitors are believed to be useful in the treatment of avariety of diseases or conditions related to systemic or tissueinflammation, inflammatory responses to infection or hypoxia, cellularactivation and proliferation, lipid metabolism, fibrosis and in theprevention and treatment of viral infections.

Bromodomain inhibitors may be useful in the treatment of a wide varietyof chronic autoimmune and/or inflammatory conditions such as rheumatoidarthritis, osteoarthritis, acute gout, psoriasis, systemic lupuserythematosus, multiple sclerosis, inflammatory bowel disease (Crohn'sdisease and Ulcerative colitis), asthma, chronic obstructive airwaysdisease, pneumonitis, myocarditis, pericarditis, myositis, eczema,dermatitis (including atopic dermatitis), alopecia, vitiligo, bullousskin diseases, nephritis, vasculitis, atherosclerosis, Alzheimer'sdisease, depression, Sjögren's syndrome, sialoadenitis, central retinalvein occlusion, branched retinal vein occlusion, Irvine-Gass syndrome(post cataract and post-surgical), retinitis pigmentosa, pars planitis,birdshot retinochoroidopathy, epiretinal membrane, cystic macular edema,parafoveal telengiectasis, tractional maculopathies, vitreomaculartraction syndromes, retinal detachment, neuroretinitis, idiopathicmacular edema, retinitis, dry eye (kerartoconjunctivitis Sicca), vernalkeratoconjunctivitis, atopic keratoconjunctivitis, anterior uveitis, panuveitis, posterior uveits, uveitis-associated macula edema. scleritis,diabetic retinopathy, diabetic macula edema, age-related maculadystrophy, hepatitis, pancreatitis, primary biliary cirrhosis,sclerosing cholangitis, Addison's disease, hypophysitis, thyroiditis,type I diabetes and acute rejection of transplanted organs.

Bromodomain inhibitors may be useful in the treatment of a wide varietyof acute inflammatory conditions such as acute gout, giant cellarteritis, nephritis including lupus nephritis, vasculitis with organinvolvement such as glomerulonephritis, vasculitis including giant cellarteritis, Wegener's granulomatosis, Polyarteritis nodosa, Behcet'sdisease, Kawasaki disease, Takayasu's Arteritis, pyoderma gangrenosum,asculitis with organ involvement and acute rejection of transplantedorgans.

Bromodomain inhibitors may be useful in the prevention or treatment ofdiseases or conditions which involve inflammatory responses toinfections with bacteria, viruses, fungi, parasites or their toxins,such as sepsis, sepsis syndrome, septic shock, endotoxaemia, systemicinflammatory response syndrome (SIRS), multi-organ dysfunction syndrome,toxic shock syndrome, acute lung injury, ARDS (adult respiratorydistress syndrome), acute renal failure, fulminant hepatitis, burns,acute pancreatitis, post-surgical syndromes, sarcoidosis, Herxheimerreactions, encephalitis, myelitis, meningitis, malaria and SIRSassociated with viral infections such as influenza, herpes zoster,herpes simplex and coronavirus.

Bromodomain inhibitors may be useful in the prevention or treatment ofconditions associated with ischaemia-reperfusion injury such asmyocardial infarction, cerebro-vascular ischaemia (stroke), acutecoronary syndromes, renal reperfusion injury, organ transplantation,coronary artery bypass grafting, cardio-pulmonary bypass procedures,pulmonary, renal, hepatic, gastro-intestinal or peripheral limbembolism.

Bromodomain inhibitors may be useful in the treatment of disorders oflipid metabolism via the regulation of APO-A1 such ashypercholesterolemia, atherosclerosis and Alzheimer's disease.

Bromodomain inhibitors may be useful in the treatment of fibroticconditions such as idiopathic pulmonary fibrosis, renal fibrosis,post-operative stricture, keloid formation, scleroderma (includingmorphea) and cardiac fibrosis.

Bromodomain inhibitors may be useful in the prevention and treatment ofviral infections such as herpes virus, human papilloma virus, adenovirusand poxvirus and other DNA viruses.

Bromodomain inhibitors may be useful in the treatment of cancer,including hematological (such as leukaemia, lymphoma and multiplemyeloma), epithelial (including lung, breast and colon carcinomas),midline carcinomas, mesenchymal, hepatic, renal and neurologicaltumours.

Bromodomain inhibitors may be useful in the treatment of dermalpathology such as non-malignant melanoma (actinic keratosis and basalcell), in-situ melanoma, squamous cell carcinoma and cutaneous T-celllymphoma.

In one embodiment the disease or condition for which a bromodomaininhibitor is indicated is selected from diseases associated withsystemic inflammatory response syndrome, such as sepsis, burns,pancreatitis, major trauma, haemorrhage and ischaemia. In thisembodiment the bromodomain inhibitor would be administered at the pointof diagnosis to reduce the incidence of: SIRS, the onset of shock,multi-organ dysfunction syndrome, which includes the onset of acute lunginjury, ARDS, acute renal, hepatic, cardiac and gastro-intestinal injuryand mortality. In another embodiment the bromodomain inhibitor would beadministered prior to surgical or other procedures associated with ahigh risk of sepsis, haemorrhage, extensive tissue damage, SIRS or MODS(multiple organ dysfunction syndrome). In a particular embodiment thedisease or condition for which a bromodomain inhibitor is indicated issepsis, sepsis syndrome, septic shock or endotoxaemia. In anotherembodiment, the bromodomain inhibitor is indicated for the treatment ofacute or chronic pancreatitis. In another embodiment the bromodomain isindicated for the treatment of burns.

In one embodiment the disease or condition for which a bromodomaininhibitor is indicated is selected from herpes simplex infections andreactivations, cold sores, herpes zoster infections and reactivations,chickenpox, shingles, human papilloma virus, human immunodeficiencyvirus (HIV), cervical neoplasia, adenovirus infections, including acuterespiratory disease, poxvirus infections such as cowpox and smallpox andAfrican swine fever virus. In one particular embodiment a bromodomaininhibitor is indicated for the treatment of Human papilloma virusinfections of skin or cervical epithelia.

The term “diseases or conditions for which a bromodomain inhibitor isindicated”, is intended to include any of or all of the above diseasestates.

In one embodiment, there is provided a method for inhibiting abromodomain which comprises contacting the bromodomain with a compoundof formula (I) or a pharmaceutically acceptable salt thereof.

While it is possible that for use in therapy, a compound of formula (I)as well as pharmaceutically acceptable salts thereof may be administeredas the raw chemical, it is common to present the active ingredient as apharmaceutical composition.

The present invention therefore provides in a further aspect apharmaceutical composition comprising a compound of formula (I) or apharmaceutically acceptable salt and one or more pharmaceuticallyacceptable carriers, diluents or excipients. The compounds of theformula (I) and pharmaceutically acceptable salts, are as describedabove. The carrier(s), diluent(s) or excipient(s) must be acceptable inthe sense of being compatible with the other ingredients of thecomposition and not deleterious to the recipient thereof. In accordancewith another aspect of the invention there is also provided a processfor the preparation of a pharmaceutical composition including admixing acompound of formula (I), or a pharmaceutically acceptable salt thereof,with one or more pharmaceutically acceptable carriers, diluents orexcipients. The pharmaceutical composition can be for use in thetreatment of any of the conditions described herein.

Since the compounds of formula (I) and pharmaceutically acceptable saltsthereof are intended for use in pharmaceutical compositions it will bereadily understood that they are each preferably provided insubstantially pure form, for example, at least 60% pure, more suitablyat least 75% pure and preferably at least 85% pure, especially at least98% pure (% in a weight for weight basis).

Pharmaceutical compositions may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Preferred unit dosage compositions are those containing a daily dose orsub-dose, or an appropriate fraction thereof, of an active ingredient.Such unit doses may therefore be administered more than once a day.Preferred unit dosage compositions are those containing a daily dose orsub-dose (for administration more than once a day), as herein aboverecited, or an appropriate fraction thereof, of an active ingredient.

Pharmaceutical compositions may be adapted for administration by anyappropriate route, for example by the oral (including buccal orsublingual), rectal, inhaled, intranasal, topical (including buccal,sublingual or transdermal), ocular (including topical, intraocular,subconjunctival, episcleral or sub-Tenon), vaginal or parenteral(including subcutaneous, intramuscular, intravenous or intradermal)route. Such compositions may be prepared by any method known in the artof pharmacy, for example by bringing into association the activeingredient with the carrier(s) or excipient(s).

In one embodiment the pharmaceutical composition is adapted forparenteral administration, particularly intravenous administration.

In one embodiment the pharmaceutical composition is adapted for oraladministration.

In one embodiment the pharmaceutical composition is adapted for topicaladministration.

A preferred dosage form that results in occlusion and modification ofskin permeation either to increase or decrease the systemic exposure ofbromodomain compounds, including but not limited to the pharmaceuticallyacceptable forms of carboxymethylcellulose, an aliginate, gelatin, orpolyvinyl pyrrolidone.

Pharmaceutical compositions adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe composition isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The compositions may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets.

Pharmaceutical compositions adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilliquid emulsions.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water and the like. Powders suitable for incorporating intotablets or capsules may be prepared by reducing the compound to asuitable fine size (e.g. by micronisation) and mixing with a similarlyprepared pharmaceutical carrier such as an edible carbohydrate, forexample, starch or mannitol. Flavoring, preservative, dispersing andcoloring agent can also be present.

Capsules may be made by preparing a powder mixture, as described above,and filling formed gelatin sheaths. Glidants and lubricants such ascolloidal silica, talc, magnesium stearate, calcium stearate or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, glidants,lubricants, sweetening agents, flavours, disintegrating agents andcoloring agents can also be incorporated into the mixture. Suitablebinders include starch, gelatin, natural sugars such as glucose orbeta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes and the like. Lubricants used in these dosageforms include sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride and the like. Disintegratorsinclude, without limitation, starch, methyl cellulose, agar, bentonite,xanthan gum and the like. Tablets are formulated, for example, bypreparing a powder mixture, granulating or slugging, adding a lubricantand disintegrant and pressing into tablets. A powder mixture is preparedby mixing the compound, suitably comminuted, with a diluent or base asdescribed above, and optionally, with a binder such ascarboxymethylcellulose, an aliginate, gelatin, or polyvinyl pyrrolidone,a solution retardant such as paraffin, a resorption accelerator such asa quaternary salt and/or an absorption agent such as bentonite, kaolinor dicalcium phosphate. The powder mixture can be granulated by wettingwith a binder such as syrup, starch paste, acadia mucilage or solutionsof cellulosic or polymeric materials and forcing through a screen. As analternative to granulating, the powder mixture can be run through thetablet machine and the result is imperfectly formed slugs broken intogranules. The granules can be lubricated to prevent sticking to thetablet forming dies by means of the addition of stearic acid, a stearatesalt, talc or mineral oil. The lubricated mixture is then compressedinto tablets. The compounds of formula (I) and pharmaceuticallyacceptable salts thereof can also be combined with a free flowing inertcarrier and compressed into tablets directly without going through thegranulating or slugging steps. A clear or opaque protective coatingconsisting of a sealing coat of shellac, a coating of sugar or polymericmaterial and a polish coating of wax can be provided. Dyestuffs can beadded to these coatings to distinguish different unit dosages.

Oral fluids such as solution, syrups and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Syrups can be prepared by dissolving thecompound in a suitably flavored aqueous solution, while elixirs areprepared through the use of a non-toxic alcoholic vehicle. Suspensionscan be formulated by dispersing the compound in a non-toxic vehicle.Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols andpolyoxy ethylene sorbitol ethers, preservatives, flavor additive such aspeppermint oil or natural sweeteners or saccharin or other artificialsweeteners, and the like can also be added.

Where appropriate, dosage unit compositions for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof can also be administered in the form of liposome deliverysystems, such as small unilamellar vesicles, large unilamellar vesiclesand multilamellar vesicles. Liposomes can be formed from a variety ofphospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

Pharmaceutical compositions adapted for topical administration may beformulated as ointments, creams, suspensions, emulsions, lotions,powders, solutions, pastes, gels, foams, sprays, aerosols or oils. Suchpharmaceutical compositions may include conventional additives whichinclude, but are not limited to, preservatives, solvents to assist drugpenetration, co-solvents, emollients, propellants, viscosity modifyingagents (gelling agents), surfactants and carriers. In one embodimentthere is provided a pharmaceutical composition adapted for topicaladministration which comprises between 0.01-10%, or between 0.01-1% ofthe compound of formula (I), or a pharmaceutically acceptable saltthereof, by weight of the composition.

For treatments of the eye or other external tissues, for example mouthand skin, the compositions are preferably applied as a topical solution,suspension, emulsion, ointment, cream, gel spray or foam. Whenformulated in an ointment, the active ingredient may be employed witheither a paraffinic or a water-miscible ointment base. Alternatively,the active ingredient may be formulated in a cream with an oil-in-watercream base or a water-in-oil. When formulated in a foam, the activeagent may be formulated with propellants, surfactants, solvents,co-solvents and viscosity modifying agents.

Pharmaceutical compositions adapted for topical administrations to theeye include eye drops wherein the active ingredient is dissolved orsuspended in a suitable carrier, especially an aqueous solvent.Compositions to be administered to the eye will have ophthalmicallycompatible pH and osmolality. One or more ophthalmically acceptable pHadjusting agents and/or buffering agents can be included in acomposition of the invention, including acids such as acetic, boric,citric, lactic, phosphoric and hydrochloric acids; bases such as sodiumhydroxide, sodium phosphate, sodium borate, sodium citrate, sodiumacetate, and sodium lactate; and buffers such as citrate/dextrose,sodium bicarbonate and ammonium chloride. Such acids, bases, and bufferscan be included in an amount required to maintain pH of the compositionin an ophthalmically acceptable range. One or more ophthalmicallyacceptable salts can be included in the composition in an amountsufficient to bring osmolality of the composition into an ophthalmicallyacceptable range. Such salts include those having sodium, potassium orammonium cations and chloride, citrate, ascorbate, borate, phosphate,bicarbonate, sulfate, thiosulfate or bisulfite anions.

An ocular delivery device may be designed for the controlled release ofone or more therapeutic agents with multiple defined release rates andsustained dose kinetics and permeability. Controlled release may beobtained through the design of polymeric matrices incorporatingdifferent choices and properties of biodegradable/bioerodable polymers(e.g. poly(ethylene vinyl) acetate (EVA), superhydrolyzed PVA),hydroxyalkyl cellulose (HPC), methylcellulose (MC), hydroxypropyl methylcellulose (HPMC), polycaprolactone, poly(glycolic) acid, poly(lactic)acid, polyanhydride, of polymer molecular weights, polymercrystallinity, copolymer ratios, processing conditions, surface finish,geometry, excipient addition and polymeric coatings that will enhancedrug diffusion, erosion, dissolution and osmosis.

Pharmaceutical compositions for ocular delivery also include in situgellable aqueous composition. Such a composition comprises a gellingagent in a concentration effective to promote gelling upon contact withthe eye or with lacrimal fluid. Suitable gelling agents include but arenot limited to thermosetting polymers. The term “in situ gellable” asused herein is includes not only liquids of low viscosity that form gelsupon contact with the eye or with lacrimal fluid, but also includes moreviscous liquids such as semi-fluid and thixotropic gels that exhibitsubstantially increased viscosity or gel stiffness upon administrationto the eye. See, for example, Ludwig (2005) Adv. Drug Deliv. Rev. 3;57:1595-639, herein incorporated by reference for purposes of itsteachings of examples of polymers for use in ocular drug delivery.

Dosage forms for nasal or inhaled administration may conveniently beformulated as aerosols, solutions, suspensions, gels or dry powders.

For compositions suitable and/or adapted for inhaled administration, itis preferred that the compound of formula (I) and pharmaceuticallyacceptable salts thereof are in a particle-size-reduced form e.g.obtained by micronisation. The preferable particle size of thesize-reduced (e.g. micronised) compound or salt is defined by a D50value of about 0.5 to about 10 microns (for example as measured usinglaser diffraction).

Aerosol formulations, e.g. for inhaled administration, can comprise asolution or fine suspension of the active substance in apharmaceutically acceptable aqueous or non-aqueous solvent. Aerosolformulations can be presented in single or multidose quantities insterile form in a sealed container, which can take the form of acartridge or refill for use with an atomising device or inhaler.Alternatively the sealed container may be a unitary dispensing devicesuch as a single dose nasal inhaler or an aerosol dispenser fitted witha metering valve (metered dose inhaler) which is intended for disposalonce the contents of the container have been exhausted.

Where the dosage form comprises an aerosol dispenser, it preferablycontains a suitable propellant under pressure such as compressed air,carbon dioxide or an organic propellant such as a hydrofluorocarbon(HFC). Suitable HFC propellants include 1,1,1,2,3,3,3-heptafluoropropaneand 1,1,1,2-tetrafluoroethane. The aerosol dosage forms can also takethe form of a pump-atomiser. The pressurised aerosol may contain asolution or a suspension of the active compound. This may require theincorporation of additional excipients e.g. co-solvents and/orsurfactants to improve the dispersion characteristics and homogeneity ofsuspension formulations. Solution formulations may also require theaddition of co-solvents such as ethanol.

For pharmaceutical compositions suitable and/or adapted for inhaledadministration, the pharmaceutical composition may be a dry powderinhalable composition. Such a composition can comprise a powder basesuch as lactose, glucose, trehalose, mannitol or starch, the compound offormula (I) or a pharmaceutically acceptable salt thereof (preferably inparticle-size-reduced form, e.g. in micronised form), and optionally aperformance modifier such as L-leucine or another amino acid and/ormetals salts of stearic acid such as magnesium or calcium stearate.Preferably, the dry powder inhalable composition comprises a dry powderblend of lactose e.g. lactose monohydrate and the compound of formula(I) or salt thereof. Such compositions can be administered to thepatient using a suitable device such as the DISKUS® device, marketed byGlaxoSmithKline which is for example described in GB 2242134 A.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof may be formulated as a fluid formulation for delivery from afluid dispenser, for example a fluid dispenser having a dispensingnozzle or dispensing orifice through which a metered dose of the fluidformulation is dispensed upon the application of a user-applied force toa pump mechanism of the fluid dispenser. Such fluid dispensers aregenerally provided with a reservoir of multiple metered doses of thefluid formulation, the doses being dispensable upon sequential pumpactuations. The dispensing nozzle or orifice may be configured forinsertion into the nostrils of the user for spray dispensing of thefluid formulation into the nasal cavity. A fluid dispenser of theaforementioned type is described and illustrated in WO-A-2005/044354.

A therapeutically effective amount of a compound of formula (I) or apharmaceutically acceptable salt thereof will depend upon a number offactors including, for example, the age and weight of the animal, theprecise condition requiring treatment and its severity, the nature ofthe formulation, and the route of administration, and will ultimately beat the discretion of the attendant physician or veterinarian. In thepharmaceutical composition, each dosage unit for oral or parenteraladministration preferably contains from 0.01 to 3000 mg, more preferably0.5 to 1000 mg, of a compound of the invention calculated as the freebase. Each dosage unit for nasal or inhaled administration preferablycontains from 0.001 to 50 mg, more preferably 0.01 to 5 mg, of acompound of the formula (I) or a pharmaceutically acceptable saltthereof, calculated as the free base.

The pharmaceutically acceptable compounds of formula (I) or apharmaceutically acceptable salt thereof can be administered in a dailydose (for an adult patient) of, for example, an oral or parenteral doseof 0.01 mg to 3000 mg per day or 0.5 to 1000 mg per day, or a nasal orinhaled dose of 0.001 to 50 mg per day or 0.01 to 5 mg per day, of thecompound of the formula (I) or a pharmaceutically acceptable saltthereof, calculated as the free base. This amount may be given in asingle dose per day or more usually in a number (such as two, three,four, five or six) of sub-doses per day such that the total daily doseis the same. An effective amount of a salt thereof, may be determined asa proportion of the effective amount of the compound of formula (I) perse.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof may be employed alone or in combination with other therapeuticagents. Combination therapies according to the present invention thuscomprise the administration of at least one compound of formula (I) or apharmaceutically acceptable salt thereof, and the use of at least oneother pharmaceutically active agent. Preferably, combination therapiesaccording to the present invention comprise the administration of atleast one compound of formula (I) or a pharmaceutically acceptable saltthereof, and at least one other pharmaceutically active agent. Thecompound(s) of formula (I) and pharmaceutically acceptable salts thereofand the other pharmaceutically active agent(s) may be administeredtogether in a single pharmaceutical composition or separately and, whenadministered separately this may occur simultaneously or sequentially inany order. The amounts of the compound(s) of formula (I) andpharmaceutically acceptable salts thereof and the other pharmaceuticallyactive agent(s) and the relative timings of administration will beselected in order to achieve the desired combined therapeutic effect.Thus in a further aspect, there is provided a combination pharmaceuticalproduct comprising a compound of formula (I) or a pharmaceuticallyacceptable salt thereof and at least one other pharmaceutically activeagent.

Thus in one aspect, the compound of formula (I) or a pharmaceuticallyacceptable salt thereof and pharmaceutical compositions comprising acompound of formula (I) or a pharmaceutically acceptable salt thereofmay be used in combination with or include one or more other therapeuticagents, for example selected from antibiotics, anti-virals,glucocorticosteroids, muscarinic antagonists, beta-2 agonists andvitamin D3 analogues. In a further aspect a compound of formula (I) or apharmaceutically acceptable salt thereof, according to the invention maybe used in combination with a further therapeutic agent which issuitable for the treatment of cancer.

It will be appreciated that when the compound of formula (I) or apharmaceutically acceptable salt thereof is administered in combinationwith other therapeutic agents normally administered by the inhaled,intravenous, oral or intranasal route, that the resultant pharmaceuticalcomposition may be administered by the same routes. Alternatively theindividual components of the composition may be administered bydifferent routes.

One embodiment of the invention encompasses combinations comprising oneor two other therapeutic agents.

It will be clear to a person skilled in the art that, where appropriate,the other therapeutic ingredient(s) may be used in the form of salts,for example as alkali metal or amine salts or as acid addition salts, orprodrugs, or as esters, for example lower alkyl esters, or as solvates,for example hydrates, to optimise the activity and/or stability and/orphysical characteristics, such as solubility, of the therapeuticingredient. It will be clear also that, where appropriate, thetherapeutic ingredients may be used in optically pure form.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical composition and thus pharmaceuticalcompositions comprising a combination as defined above together with apharmaceutically acceptable diluent or carrier represent a furtheraspect of the invention.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof may be prepared by the methods described below or by similarmethods. Thus the following Intermediates and Examples serve toillustrate the preparation of the compounds of formula (I) andpharmaceutically acceptable salts thereof, and are not to be consideredas limiting the scope of the invention in any way.

GENERAL EXPERIMENTAL DETAILS

All temperatures referred to are in ° C.

The names of the following compounds have been obtained using thecompound naming programme “ACD Name Pro 6.02” or Chem Draw Ultra 12.0.

Abbreviations

AcOH refers to acetic acid

BINAP refers to 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl

BOC refers to tert-butoxycarbonyl

CV refers to column volumes

DCM refers to dichloromethane

1,2-DCE refers to 1,2-dichloroethane

DIPEA refers to diisopropylethylamine

DMSO refers to dimethylsulfoxide.

DMF refers to N,N-dimethylformamide

Ether refers to diethyl ether

Et₂O refers to diethyl ether

EtOAc refers to ethyl acetate

FMOC refers to 9-fluorenylmethoxycarbonyl

HATU refers to O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate

HPLC refers to high performance liquid chromatography

IPA refers to propan-2-ol

i-Pr₂O refers to di-isopropyl ether

LiAlH₄ refers to lithium aluminium hydride

MDAP refers to Mass directed autoprep refers preparative mass directedHPLC

MeCN refers to acetonitrile

MeOH refers to methanol

MgSO₄ refers to magnesium sulfate

Mp refers to melting point

r.t. refers to room temperature

Rt refers to retention time

Na₂SO₄ refers to sodium sulphate

TBTU refers to O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumtetrafluoroborate

TMEDA refers to tetramethylethylenediamine

TFA refers to trifluoroacetic acid

THF refers to tetrahydrofuran

TLC refers to thin layer chromatography

LCMS Methodology

Method Formate (Formic Acid Modifier)

LC Conditions

The UPLC analysis was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm, i.d. 1.7 μm packing diameter) at 40° C.

The solvents employed were:

-   -   A=0.1% v/v solution of formic acid in water    -   B=0.1% v/v solution of formic acid in acetonitrile

The gradient employed was:

Time (min) Flow rate (ml/min) % A % B 0 1 99 1 1.5 1 3 97 1.9 1 3 97 2.01 0 100

The UV detection was a summed signal from wavelength of 210 nm to 350nm.

MS Conditions

MS: Waters ZQ

Ionisation mode: Alternate-scan positive and negative electrospray

Scan range: 100 to 1000 AMU

Scan time: 0.27 sec

Inter scan delay: 0.10 sec

Method HpH (Ammonium Bicarbonate Modifier)

LC Conditions

The UPLC analysis was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm, i.d. 1.7 μm packing diameter) at 40° C.

The solvents employed were:

-   -   A=10 mM ammonium hydrogen carbonate in water adjusted to pH10        with ammonia solution    -   B=acetonitrile

The gradient employed was:

Time (min) Flow rate (ml/min) % A % B 0 1 99 1 1.5 1 3 97 1.9 1 3 97 2.01 0 100

The UV detection was a summed signal from wavelength of 210 nm to 350nm.

MS Conditions

MS: Waters ZQ

Ionisation mode: Alternate-scan positive and negative electrospray

Scan range: 100 to 1000 AMU

Scan time: 0.27 sec

Inter scan delay: 0.10 sec

MDAP Methodology

Method Formate (Formic Acid Modifier)

LC Conditions

The HPLC analysis was conducted on either a Sunfire C18 column (100mm×19 mm, i.d 5 μm packing diameter) or a Sunfire C18 column (150 mm×30mm, i.d. 5 μm packing diameter) at ambient temperature.

The solvents employed were:

-   -   A=0.1% v/v solution of formic acid in water    -   B=0.1% v/v solution of formic acid in acetonitrile

Run as a gradient over either 15 or 25 min (extended run) with a flowrate of 20 ml/min (100 mm×19 mm, i.d 5 μm packing diameter) or 40 ml/min(150 mm×30 mm, i.d. 5 μm packing diameter).

The UV detection was a summed signal from wavelength of 210 nm to 350nm.

MS Conditions

MS: Waters ZQ

Ionisation mode: Alternate-scan positive and negative electrospray

Scan range: 100 to 1000 AMU

Scan time: 0.50 sec

Inter scan delay: 0.20 sec

Method HpH (Ammonium Bicarbonate Modifier)

LC Conditions

The HPLC analysis was conducted on either an Xbridge C18 column (100mm×19 mm, i.d 5 μm packing diameter) or a Xbridge C18 column (100 mm×30mm, i.d. 5 μm packing diameter) at ambient temperature.

The solvents employed were:

-   -   A=10 mM ammonium bicarbonate in water, adjusted to pH10 with        ammonia solution    -   B=acetonitrile

Run as a gradient over either 15 or 25 min (extended run) with a flowrate of 20 ml/min (100 mm×19 mm, i.d 5 μm packing diameter) or 40 ml/min(100 mm×30 mm, i.d 5 μm packing diameter).

The UV detection was a summed signal from wavelength of 210 nm to 350nm.

MS Conditions

MS: Waters ZQ

Ionisation mode: Alternate-scan positive and negative electrospray

Scan range: 100 to 1000 AMU

Scan time: 0.50 sec

Inter scan delay: 0.20 sec

Intermediate 11-((2S,4R)-4-Amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone

A suspension of aluminium chloride (41.2 g, 309 mmol) in DCM (480 mL) at0° C. under nitrogen was treated with a solution of isopropyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(30 g, 81 mmol) in DCM (80 ml) via cannula and the resulting mixture wasstirred at this temperature for 30 min. The reaction mixture was thenslowly treated with a mixture of triethylamine (136 mL, 975 mmol) andMeOH (48 mL) via cannula. The resulting cake formed was stirred in EtOAc(800 mL), isolated by filtration and subsequently partitioned betweenDCM (800 mL) and saturated aqueous NaHCO₃ solution (800 mL). Sodiumpotassium tartrate (300 g) was added and the resulting mixture wasstirred vigorously for 2 h. The layers were separated and the DCM layerwas filtered through a sinter funnel. The filtrate was dried (MgSO₄) andconcentrated in vacuo to give a first crop of1-((2S,4R)-4-amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(19.6 g, 69.2 mmol, 85% yield) as a yellow foam. The aqueous phase wastreated further with DCM (800 mL) and the biphasic mixture stirredovernight. The layers were then separated and the organic layer wasdried over (MgSO₄) and concentrated in vacuo to give a second crop of1-((2S,4R)-4-amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(3.4 g, 12.01 mmol, 14.78% yield). LCMS (HpH, 2 min), Rt=0.77 min,MH+=283 (1 Br).

Intermediate 26-(((2S,4R)-1-Acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

To a mixture of1-((2S,4R)-4-amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 1)(2.28 g, 8.05 mmol) and6-chloronicotinonitrile (2.231 g, 16.10 mmol) was added NMP (20 mL) andthe mixture treated with DIPEA (4.22 mL, 24.16 mmol). The mixture wassplit between 2 flasks and each flask was flushed with nitrogen, sealedand stirred under microwave irradiation at 200° C. for 2 h. The reactionmixtures were combined and partitioned between water and EtOAc. Theaqueous layer was extracted with EtOAc (×4) and the combined organiclayers were washed with water (×3), then brine and were dried (MgSO₄)filtered and concentrated in vacuo. The brown solid residue was purifiedby chromatography (EtOAc in Hexanes gradient) to give6-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(1.940 g, 5.04 mmol, 62.5% yield) as a pale yellow foam. LCMS (HpH, 2min), Rt=1.02 min, MH+=386 (1 Br).

Intermediate 3 Methyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a flask charged with6-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 2)(1000 mg, 2.60 mmol),(4-(methoxycarbonyl)phenyl)boronic acid (561 mg, 3.11 mmol),tetrakis(triphenylphosphine)palladium(0) (300 mg, 0.260 mmol) andpotassium carbonate (1076 mg, 7.79 mmol) was added DME (20 mL) and water(4.0 mL). The resulting mixture was stirred at 100° C. under nitrogenfor 1 h, then cooled to room temperature and concentrated in vacuo. Theresidue was partitioned between EtOAc and water and the layers wereseparated. The organic phase was dried (MgSO₄), concentrated in vacuoand the residue was purified by chromatography (10 g column, MeOH/DCMgradient) to give methyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(1002 mg, 2.275 mmol, 88% yield) as an orange foam.

LCMS (HpH, 2 min), Rt=1.09 min, MH+=441.

Intermediate 44-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid

A solution of methyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 3) (1.0 g, 2.270 mmol) in MeOH (15mL) at room temperature was treated with aqueous sodium hydroxidesolution (2N, 2.27 mL, 4.54 mmol) and the resulting mixture was stirredat this temperature for 24 h. The bulk of MeOH was evaporated in vacuoand the aqueous residue was treated with acetic acid (0.39 mL, 6.81mmol) giving precipitate which was isolated by filtration and driedunder vacuum at 40° C. to afford4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (820 mg, 1.923 mmol, 85% yield) as a yellow solid. LCMS (HpH, 2min), Rt=0.64 min, MH+=427.

Intermediate 56-(((2S,4R)-1-Acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

To a flask charged with6-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 2) (847 mg, 2.199 mmol),bis(pinacolato)diboron (1228 mg, 4.84 mmol), PdCl₂(dppf) (161 mg, 0.220mmol) and potassium acetate (324 mg, 3.30 mmol) was added DMSO (7 mL),the mixture degassed under nitrogen and stirred under nitrogen for 1 hat 80° C. Further portions of bis(pinacolato)diboron (1 g), PdCl₂(dppf)(100 mg) and potassium acetate (150 mg) were added and the mixture wasstirred for 45 min. Further portions of bis(pinacolato)diboron (1 g),PdCl₂(dppf) (100 mg) and potassium acetate (150 mg) was added and themixture was stirred for 45 min. The reaction mixture was cooled to roomtemperature and treated with EtOAc and water. The biphasic mixture wasfiltered through a pad of Celite™ (10 g) and the layers were separated.The aqueous layer was extracted with EtOAc (×2) and the combined organiclayers were washed with water (×4), then brine, dried (MgSO₄) andconcentrated in vacuo. Purification of the residue by chromatography [50g column, EtOAc/Hexanes gradient] afforded6-(((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(800 mg, 1.850 mmol, 84% yield) as a red gum. LCMS (Formate, 2 min),Rt=1.08 min, MH+=433.

Intermediate 6 Methyl6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate

To a flask charged with6-(((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 5) (800 mg, 1.850 mmol), methyl6-bromonicotinate (440 mg, 2.036 mmol), potassium carbonate (767 mg,5.55 mmol) and tetrakis(triphenylphosphine)palladium(0) (214 mg, 0.185mmol) were added toluene (8 mL) and ethanol (8 mL). The resultingmixture was stirred at 90° C. under nitrogen for 3 h at which pointportions of potassium carbonate (384 mg)tetrakis(triphenylphosphine)palladium(0) (107 mg) and methyl6-bromonicotinate (220 mg) were added and the reaction mixture stirringcontinued for 5 h. The mixture was then cooled to room temperature andconcentrated in vacuo. The residue was partitioned between EtOAc andwater. The phases were separated and the aqueous layer was extractedwith EtOAc. The combined organic layers were washed with brine, dried(MgSO₄) and concentrated in vacuo. The residue was purified bychromatography [25 g column, EtOAc/Hexanes gradient] to give ethyl6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(600 mg, 1.317 mmol, 71.2% yield) as a white foam. LCMS (HpH, 2 min),Rt=1.07 min, MH+=456.

Intermediate 76-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid

A solution of methyl6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(for a preparation see Intermediate 6) (580 mg, 1.273 mmol) in ethanol(10 mL) was treated with a aqueous lithium hydroxide solution (1N, 2.55mL, 2.55 mmol) and the resulting mixture was stirred for 2 h. The bulkof ethanol was evaporated in vacuo and the residue was diluted withwater (ca. 5 mL). The cloudy mixture was treated with acetic acid (0.146mL, 2.55 mmol) and the precipitate formed was isolated by filtration,washed with Et₂O and dried at 60° C. for 16 h to give6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (410 mg, 0.959 mmol, 75% yield) as a pale yellow solid which wasused in the next step without further purification. LCMS (HpH, 2 min),Rt=0.63 min, MH+=428.

Intermediate 8 Ethyl2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)acetate

To a solution of6-(((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 5) (1.67 g, 3.86 mmol), ethyl2-(4-bromophenyl)acetate (1.127 g, 4.64 mmol) and potassium carbonate(1.602 g, 11.59 mmol) in toluene (10 mL) and ethanol (10.0 mL) was addedtetrakis(triphenylphosphine)palladium(0) (0.446 g, 0.386 mmol) undernitrogen. The reaction mixture heated at 100° C. for 1 h, thenpartitioned between EtOAc and water. The layers were separated and theaqueous phase was extracted with EtOAc (×3). The combined organic layerswere washed with brine, dried (MgSO₄), filtered and concentrated invacuo. The residue was purified by flash chromatography on silica gel(25 g), eluting with an EtOAc/cyclohexane gradient (10 to 80%). Theappropriate fractions were combined and concentrated in vacuo to giveethyl2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)acetate(752 mg, 42%) as a viscous colourless oil. LCMS (Formate, 2 min),Rt=1.10 min, MH+=469.

Intermediate 92-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid, lithium salt

To a solution of ethyl2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)acetate(for a preparation see Intermediate 8) (300 mg, 0.640 mmol) in MeOH (5mL) was added lithium hydroxide solution (0.768 mL, 0.768 mmol). Theresulting mixture was stirred at 40° C. for 2 h, whereupon it wasconcentrated under vacuum to give the crude lithium carboxylate of2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid (286 mg, 100%) as a white solid which was used in the next stepwithout further purification. LCMS (Formate, 2 min), Rt=1.10 min,MH+=433.

Intermediate 103-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)propanoicacid

To a solution of6-(((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 5) (382 mg, 0.884 mmol) and3-(4-bromophenyl)propanoic acid (243 mg, 1.060 mmol) in toluene (6 mL)and ethanol (6 mL) were successively addedtetrakis(triphenylphosphine)palladium(0) (102 mg, 0.088 mmol) andpotassium carbonate (366 mg, 2.65 mmol). The resulting mixture wasstirred at 80° C. for 2 h, then partitioned between water and EtOAc. Thelayers were separated and the aqueous phase was extracted with EtOAc(×3). The combined organic layers were washed with brine, dried (MgSO₄),filtered and concentrated in vacuo. The crude residue was purified byflash chromatography on silica gel (25 g), eluting with EtOAc incyclohexane (10-70%). The appropriate fractions were combined andconcentrated under reduced pressure to give3-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)propanoicacid (209 mg, 52%) as a white solid. LCMS (Formate, 2 min), Rt=0.91 min,MH+=455.

Intermediate 11 1-Methylethyl (2E)-2-butenoylcarbamate

Isopropyl carbamate (30 g, 291 mmol, available from TCl) was charged toa 3 L Lara vessel and dry tetrahydrofuran (THF) (150 ml) added.(2E)-2-Butenoyl chloride (31.2 ml, 326 mmol, available from Aldrich) wasadded under nitrogen and the jacket cooled to −30° C. When the solutiontemperature reached −17° C. Lithium tert-butoxide (1M, 655 mL, 655 mmol)was added by peristaltic pump over 2 h, keeping the reaction temperaturebetween −10° C. and −18° C. Once the addition was complete the mixturewas stirred for 30 min and brought to 0° C. Diethyl ether (450 ml) andhydrochloric acid (1M, 375 mL) were added and the mixture brought to 20°C. with vigourous stirring. The stirring was stopped, the layers allowedto separate and the aqueous layer run off. Brine (375 mL) was added andthe mixture stirred vigorously. The stirring was stopped, the layersallowed to separate and the aqueous layer run off. The organic layer wasdried (MgSO₄), filtered and evaporated to a brown oil (60 g). The wasapplied to a silica column (40+M Biotage) and eluted with DCM/EtOAc (1:1to 0:1, 10CV). The product containing fractions were evaporated todryness and loaded on to a Redisep Isco silica column (1500 g) andeluted with a EtOAc in cyclohexane gradient (0-40%). The clean, productcontaining fractions were evaporated to an off-white solid (15.41 g).LCMS (Method C): Rt=0.68, MH+=172

Intermediate 12 1-Methylethyl{(3S)-3-[(4-bromophenyl)amino]butanoyl}carbamate

1-Methylethyl (2E)-2-butenoylcarbamate (for a preparation seeIntermediate 11)(9.38 g, 54.8 mmol) was stirred in toluene (281 mL)under nitrogen and (R-BINAP)ditriflatebis(acetonitrile)palladium(II)(for a preparation see Intermediate 36)(3.35 g, 3.01 mmol) added. Thecatalyst formed a gummy ball, the solution turned to an opaque yellowmixture and was stirred for 20 min. 4-Bromoaniline (14.14 g, 82 mmol)was added, the solution turned to a clear light brown and the gummycatalyst dissolved further. The mixture was stirred fior 16 h. Similarlya second batch of 1-methylethyl (2E)-2-butenoylcarbamate (Intermediate11, 8.51 g, 49.7 mmol) was stirred in toluene (255 mL) under nitrogenand (R-BINAP)ditriflatebis(acetonitrile)palladium(II) (3.04 g, 2.73mmol) added. The catalyst formed a gummy ball, the solution turned to anopaque yellow mixture and was stirred for 20 min. 4-Bromoaniline (12.83g, 74.6 mmol) was added, the solution turned to a clear light brown andthe gummy catalyst dissolved further. The mixture was stirred for 16 h.

The two reaction mixtures were combined and loaded on to a 1.5 kg Iscosilica Redisep column. The column was eluted with DCM/MeOH (0%→0.5%, 19CV). The clean, product containing fractions were evaporated to a palebrown oil. The mixture was dried in a vacuum oven overnight at 40° C. togive 1-methylethyl {(3S)-3-[(4-bromophenyl)amino]butanoyl}carbamate as awhite solid (24.2 g, 67% overall).

LCMS (Method C): Rt=0.91, MH+=343. ee=92%.

Intermediate 13 1-Methylethyl[(2S,4R)-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl]carbamate

1-Methylethyl {(3S)-3-[(4-bromophenyl)amino]butanoyl}carbamate (for apreparation see Intermediate 12) (17.9 g, 52.2 mmol) was taken up inethanol (150 mL) and cooled to below −10° C. (internal temperature) in aCO₂/acetone bath. NaBH₄ (1.381 g, 36.5 mmol) was added followed bymagnesium chloride hexahydrate (11.35 g, 55.8 mmol) in water (25 mL)keeping the temperature below −5° C. The mixture was allowed to stir at<0° C. for 1 h then warmed to room temperature and stirred for 1 h. Theresulting thick suspension was poured into a mixture of citric acid(25.05 g, 130 mmol), HCl (1M in water, 205 mL, 205 mmol) and DCM (205mL). The biphasic mixture was stirred at room temperature for 1 h. Thelayers were separated and the organic layer dried with Na₂SO₄, filteredand concentrated to yield the product as a light brown solid (14.1 g).LCMS (Method B): Rt=1.13, MH+=327

Intermediate 14 1-Methylethyl[(2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl]carbamate

1-Methylethyl[(2S,4R)-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl]carbamate (fora preparation see Intermediate 13) (14.1 g, 43.1 mmol) was taken up inDCM (400 mL) under nitrogen at room temperature. Pyridine (10.46 mL, 129mmol), then acetyl chloride (4.60 mL, 64.6 mmol), were added and thereaction stirred at room temperature for 16 h, then partitioned betweenEtOAc (2000 mL) and a saturated NaHCO₃ aqueous solution (800 mL). Thelayers were separated and the organic phase was washed with water thenbrine (1500 mL each) and then dried with Na₂SO₄ and concentrated invacuo to yield a purple solid. The crude product was taken up in theminimum of DCM and applied to a 330 g Companion XL column and elutedwith a gradient of 12-63% Ethyl Acetate in cyclohexane to give theproduct as an off-white solid (12.37 g).

LCMS (Method B): Rt=1.03, MH+=369

[alpha]D=+281.1025° (T=20.7° C., 10 mm cell, c=0.508 g/100 ml, ethanol).

Intermediate 15 Ethyl4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]benzoate

1-Methylethyl[(2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl]carbamate(for a preparation see Intermediate 14), (39.0 g, 106 mmol),{4-[(ethyloxy)carbonyl]phenyl}boronic acid (22.5 g, 116 mmol) andtetrakis(triphenylphosphine)palladium(0) (1.83 g, 1.58 mmol) were mixedin DME (430 mL) and the resulting mixture was treated with aqueousNa₂CO₃ (2N, 210 mL, 420 mmol). The mixture was degassed under housevacuum with several quenches with nitrogen and then stirred at 105° C.under nitrogen for approximately 6 h before being allowed to cool toroom temperature. The mixture was partitioned between EtOAc and waterand the layers were separated. The aqueous phase was extracted withEtOAc and the combined organic phases were washed with brine. Theorganic phase was then filtered through a 70 g silica cartridge, washingthe cartridge with EtOAc. The combined filtrate and washings wereconcentrated in vacuo. The residue was triturated with Et₂O thenfiltered off. The solid obtained was air-dried to give ethyl4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]benzoate(for a preparation see Intermediate 15) (35.2 g, 80.2 mmol, 76%) as agrey solid. The filtrate was concentrated in vacuo and the residueobtained triturated with Et₂O (approximately 30 mL). The solid formedwas isolated by filtration and air-dried, to give ethyl4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]benzoateas a grey solid (5.96 g, 13.5 mmol, 13%). LCMS (formate, 2 min),Retention time 1.16 min, MH+=439

Intermediate 16 Ethyl4-[(2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl]benzoate

Ethyl4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]benzoate(for a preparation see intermediate 15) (8.90 g, 20.30 mmol) was addedto a suspension of aluminium chloride (10.3 g, 77 mmol) in DCM (160 mL)cooled with an ice/water bath. The temperature rose from 0° C. toapproximately 6° C. after the addition. The resulting mixture wasstirred at approximately 0° C. for 20 min, and then treated with asolution of MeOH (18 mL) and triethylamine (34 mL, 245 mmol) over ˜30sec. The resulting mixture was stirred at 0° C. for ˜30 min, and thenpartitioned between EtOAc and a saturated NaHCO₃ aqueous solution.

The same reaction was done in parallel, using of ethyl4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]benzoate(for a preparation see Intermediate 15) (0.89 g 2.030 mmol), aluminiumchloride (1.03 g, 7.72 mmol), triethylamine (3.4 mL, 24.53 mmol), DCM(16 mL) and MeOH (1.3 mL). The products of both reactions were combinedat this stage and the resulting mixture was stirred at room temperaturefor approximately 10 min (total volume: approximately 1 L). The mixturewas filtered through Celite™, the insoluble residue was washed withEtOAc and a saturated NaHCO₃ aqueous solution and the layers wereseparated. The aqueous phase was extracted with EtOAc and the combinedorganic phases were washed with brine, dried (hydrophobic frit) andconcentrated in vacuo to give ethyl4-[(2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl]benzoate(6.6 g, 84%—allowing for the addition of the parallel experiment) as acream solid. LCMS (formate, 2 min), Retention time 0.73 min,[M-NH₂]+=336

Intermediate 17 Ethyl4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoate

Ethyl4-[(2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl]benzoate(for a preparation see Intermediate 16) (6.6 g, 18.73 mmol),1-bromo-4-chlorobenzene (3.94 g, 20.60 mmol),bis(dibenzylideneacetone)palladium (0) (690 mg, 1.2 mmol) and[2′-(dicyclohexylphosphanyl)-2-biphenylyl]dimethylamine (Dave-phos) (590mg, 1.499 mmol)) were mixed in toluene (120 mL) and the resultingmixture was treated with sodium t-butoxide (2.52 g, 26.2 mmol). Thereaction was degassed under house vacuum with several quenches withnitrogen, heated at 70° C. under nitrogen for 16 h, then was allowed tocool to room temperature and filtered. The insoluble residue was washedwith toluene and then Et₂O. The combined filtrate and washings werewashed with water (×2) then extracted with hydrochloric acid (2N, ×20,resulting in the precipitation of an orange oil which was collected withthe aqueous acidic phases. The acidic extracts were washed with Et₂O andthe combined organic phases were washed with brine, dried (hydrophobicfrit) and concentrated in vacuo. The residue was purified bychromatography on a silica cartridge (330 g), eluting with anEtOAc/cyclohexane gradient (5-45%). The appropriate fractions werecombined and reduced to dryness in vacuo to give a pale yellow foam.This foam was dissolved in EtOAc (50 mL) and treated with functionalthiourea silica (0.56 g, palladium scavenger). The mixture was stirredat room temperature (air atmosphere) for ˜20 min and then left at roomtemperature for 16 h. The mixture was filtered and the insolubleresidues washed with EtOAc. The combined filtrate and washings wereconcentrated in vacuo to give ethyl4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoate(3.7 g, 8.0 mmol, 32%) as a yellow oil.

Intermediate 184-{(2S,4R)-1-Acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoicacid

Ethyl4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoate(for a preparation see Intermediate 17) (5.41 g, 11.69 mmol) wasdissolved in ethanol (100 mL) and the solution was treated with aqueousNaOH solution (2M, 50 mL, 100 mmol). The resulting mixture was stirredat room temperature (air atmosphere) for approximately 2 h then most ofthe ethanol was removed in vacuo. The resulting yellow solution wasdiluted with water (resulting in the formation of an oily yellowprecipitate). The aqueous phase was washed twice with DCM (which didn'tdissolve the precipitate previously formed) then was acidified withhydrochloric acid (2N) to pH 1 and extracted with EtOAc (×2). Thecombined EtOAc phases were washed with brine, dried (hydrophobic frit)and concentrated in vacuo. The residual yellow foam was triturated withEt₂O over approximately 1 h. The resulting solid was isolated byfiltration, washed with Et₂O and air-dried to give4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoicacid (4.41 g, 10.1 mmol, 87%) as a cream solid. LCMS (HpH), Retentiontime 1.08 min, [M−H]-=433

Intermediate 191-((2S,4R)-6-bromo-2-methyl-4-((5-nitropyridin-2-yl)amino)-3,4-dihydroquinolin-1(2H)-yl)ethanone

To a flask charged with1-((2S,4R)-4-amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 1)(2 g, 7.06 mmol) and2-chloro-5-nitropyridine (2.240 g, 14.13 mmol) was added NMP (10 mL) andthe resulting mixture treated with DIPEA (3.70 mL, 21.19 mmol). Themixture was divided between 2 flasks, each flask flushed with nitrogen,sealed and the mixture stirred under microwave irradiation at 200° C.for 2 h. The 2 reaction mixtures were combined, diluted with water andextracted with EtOAc (×4). Combined EtOAc layers were washed with water(×3) then brine, dried over MgSO₄ and concentrated in vacuo. Theresidual brown solid was purified by chromatography eluting with anEtOAc/hexanes gradient, which gave1-((2S,4R)-6-bromo-2-methyl-4-((5-nitropyridin-2-yl)amino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(4.6 g, 11.35 mmol, 161% yield) as a pale brown foam. LCMS (HpH, 2 min),Rt=1.08 min, MH+=405/407 (doublet).

Intermediate 20 Methyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a flask charged with(2S,4R)-1-acetyl-6-bromo-2-methyl-N-(5-nitro-2-pyridinyl)-1,2,3,4-tetrahydro-4-quinolinamine(for a preparation see Intermediate 19) (4.1 g, 10.12 mmol),{4-[(methyloxy)carbonyl]phenyl}boronic acid (2.185 g, 12.14 mmol),tetrakis(triphenylphosphine)palladium(0) (1.169 g, 1.012 mmol) andpotassium carbonate (4.19 g, 30.4 mmol) was added DME (50 mL) and water(10.0 mL) and the resulting mixture was stirred at 100° C. undernitrogen for 1 h then cooled to room temperature and concentrated invacuo. The residue was partitioned between EtOAc and water and thelayers were separated. The organic phase was dried over MgSO4 andconcentrated in vacuo. The residue was purified by chromatographyeluting with an EtOAc/hexanes gradient, which gave methyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(4.3 g, 9.34 mmol, 92% yield) as an orange foam, LCMS (HpH, 2 min),Rt=1.13 min, MH+=461.

Intermediate 21 Methyl4-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a flask charged with methyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 20)(2.43 g, 5.28 mmol), ammoniumformate (3.33 g, 52.8 mmol) and palladium on carbon (0.562 g, 5.28 mmol)was added MeOH (50 mL) and the resulting mixture was refluxed for 30min. The reaction was cooled to room temperature, filtered throughCelite™ and the residue washed with MeOH then DMF. The methanolicfiltrate and washings were combined and concentrated in vacuo. Theresidue was dissolved in EtOAc, washed with brine, dried over MgSO4 andconcentrated in vacuo. The residue was purified by chromatographyeluting with a MeOH/DCM gradient to give methyl4-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(2.0 g, 4.65 mmol, 88% yield) as a dark pink foam. LCMS (HpH, 2 min),Rt=0.94 min, MH+=431.

Intermediate 22 Methyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

A solution of methyl4-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 21) (1.04 g, 2.416 mmol) in ethanol(20 mL) at room temperature was treated with isoamyl nitrite (0.813 mL,6.04 mmol) and the resulting mixture was stirred for 2 h at reflux thencooled to room temperature. Most of the solvent was removed in vacuo.The residue was treated with EtOAc and brine and both layers were passedthrough 10G of Celite™ then separated. The organic layer was dried overMgSO₄ and concentrated in vacuo.

The residue was purified by chromatography, eluting with a MeOH/DCMgradient, which gave methyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(720 mg, 1.733 mmol, 71.7% yield) as a light pink foam. LCMS (HpH, 2min), Rt=1.10 min, MH+=416.

Intermediate 234-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid

Methyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 22)(700 mg, 1.685 mmol) wasdissolved in MeOH (10 ml) and sodium hydroxide solution (2M, 1.68 ml,3.37 mmol) was added. After 6.5 h a further 2 equivalents of sodiumhydroxide (2M, 1.685 ml) was added and reaction mixture stirredovernight. The was concentrated and the residue was diluted with water.Hydrochloric acid (2N, 1 ml) was added forming a precipitate which wasisolated by filtration and dried to give4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (500 mg, 1.245 mmol, 73.9% yield) as a pinky solid. The aqueousphase was extracted EtOAc (×3) and the combined organic phases weredried over MgSO₄ then concentrated in vacuo to give a further batch of4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (140 mg, 0.349 mmol, 20.70% yield) as a yellow foam. LCMS (HpH, 2min), Rt=0.62 min, MH+=402.

Intermediate 241-((2S,4R)-2-methyl-4-((5-nitropyridin-2-yl)amino)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

To a flask charged with1-((2S,4R)-6-bromo-2-methyl-4-((5-nitropyridin-2-yl)amino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 19)(1.17 g, 2.89 mmol),bis(pinacolato)diboron (1.613 g, 6.35 mmol), PdCl₂(dppf) (0.211 g, 0.289mmol) and potassium acetate (0.425 g, 4.33 mmol) was added DMSO (10 mL).The reaction mixture was degassed (under house vacuum over 10 min withseveral quenches with nitrogen) and stirred under nitrogen for 1.5 h at80° C. The reaction was cooled to room temperature over 2 h, and dilutedwith EtOAc and water. The biphasic mixture was filtered through a pad ofCelite™ (10 g) and the layers were separated. The aqueous layer wasextracted with EtOAc (×2) and the combined organic phases were washedwith water (×4) then brine, dried over MgSO₄ and concentrated in vacuo.The residue was purified by chromatography eluting with an EtOAc/Hexanesgradient; which gave1-((2S,4R)-2-methyl-4-((5-nitropyridin-2-yl)amino)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(1.03 g, 2.277 mmol, 79% yield) as a light brown foam. LCMS (HpH, 2min), Rt=1.15 min, MH+=453.

Intermediate 25 ethyl6-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate

to a flask charged with1-((2S,4R)-2-methyl-4-((5-nitropyridin-2-yl)amino)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 24)(1.03 g, 2.277 mmol), methyl6-bromonicotinate (0.541 g, 2.505 mmol), potassium carbonate (0.944 g,6.83 mmol) and tetrakis(triphenylphosphine)palladium (0) (0.263 g, 0.228mmol) was added a previously degassed mixture of toluene (10 mL) andethanol (10 mL). The resulting mixture was stirred at 90° C. undernitrogen. After 3 h, further portions of methyl 6-bromonicotinate (0.271g, 1.252 mmol), potassium carbonate (0.472 g, 3.42 mmol) andtetrakis(triphenylphosphine)palladium (0) (0.132 g, 0.114 mmol) wereadded and reaction mixture heated for a further 4 h. The reaction wascooled to room temperature and concentrated in vacuo. The crude residuewas partitioned between EtOAc and water, the phases separated andaqueous phase extracted with EtOAc. The combined organic phases werewashed with brine, dried over MgSO₄ and concentrated in vacuo.Purification of the residue by chromatography, eluting with anEtOAc/hexanes gradient gave ethyl6-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(740 mg, 1.556 mmol, 68.3% yield) as a yellow foam.

LCMS (HpH, 2 min), Rt=1.15 min, MH+=476.

Intermediate 26 ethyl6-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate

To a flask charged with ethyl6-((2S,4R)-1-acetyl-2-methyl-4-((5-nitropyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(for a preparation see Intermediate 25)(780 mg, 1.640 mmol), ammoniumformate (1034 mg, 16.40 mmol) and palladium on carbon (10% Pd/C) (160mg, 1.503 mmol) was added ethanol (15 ml) and the resulting mixture wasrefluxed for 45 min. The reaction was cooled to room temperature,filtered through Celite™, the residue washed with EtOH and the combinedfiltrate and washings concentrated in vacuo. The residue was dissolvedin EtOAc, washed with brine, dried over MgSO₄ and concentrated in vacuo.The residue was purified by chromatography eluting with a MeOH/DCMgradient; which gave ethyl6-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(450 mg, 1.010 mmol, 61.6% yield) as a pink foam. LCMS (HpH, 2 min),Rt=0.93 min, MH+=446.

Intermediate 27 ethyl6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate

A solution of ethyl6-((2S,4R)-1-acetyl-4-((5-aminopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(for a preparation see Intermediate 26)(450 mg, 1.010 mmol) in ethanol(10 mL) at room temperature was treated with isoamyl nitrite (0.34 mL,2.53 mmol) and the resulting mixture was stirred 1 h at reflux thencooled to room temperature. Most of the solvent was evaporated in vacuo.The residue was treated with EtOAc and brine and both layers werefiltered through Celite™ (10 g) then separated. The organic layer wasdried over MgSO₄ and concentrated in vacuo.

The residue was purified by chromatography, eluting with a MeOH/DCMgradient; which gave ethyl6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(240 mg, 0.557 mmol, 55.2% yield) as a light pink foam. LCMS (HpH, 2min), Rt=1.09 min, MH+=431.

Intermediate 286-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid

A solution of ethyl6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(for a preparation see Intermediate 27)(240 mg, 0.557 mmol) in ethanol(5 mL) was treated with aqueous lithium hydroxide (1N, 1.115 mL, 1.115mmol) and the resulting mixture was stirred at room temperature. After 1h most of the ethanol was removed in vacuo, the residue was diluted withwater (ca 2 mL) and the “cloudy” mixture was treated with acetic acid(0.064 mL, 1.115 mmol). The precipitate formed was isolated byfiltration, washed with Et₂O and dried at 60° C. for 16 h to give6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinic acid (150 mg, 0.373 mmol, 66.9% yield) as a pale yellow solid whichwas used in the next step without further purification. LCMS (HpH, 2min), Rt=0.61 min, MH+=403.

Intermediate 29 tert-butyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

A solution of(2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinamine(11.3 g, 39.9 mmol) in DCM (120 mL) at room temperature was treated withtriethylamine (16.69 mL, 120 mmol) then Boc₂O (12.97 mL, 55.9 mmol) andthe resulting mixture was stirred at this temperature for 1 h. Furtherportions of base and anhydride (20%) were added further and after 30min, the mixture was washed with saturated aqueous NaHCO₃ solution,dried (hydrophobic frit) and concentrated in vacuo. The residue waspurified by chromatography, eluting with an EtOAc/hexane gradient; whichgave tert-butyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(9.2 g, 24.00 mmol, 60.1% yield) as a white foam. LCMS (HpH, 2 min),Rt=1.14 min, MH+=383/385.

Intermediate 30 methyl4-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a flask charged with tert-butyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation see Intermediate 29)(5 g, 13.05 mmol),(4-(methoxycarbonyl)phenyl)boronic acid (2.58 g, 14.35 mmol),tetrakis(triphenylphosphine)palladium (0) (1.507 g, 1.305 mmol) andpotassium carbonate (5.41 g, 39.1 mmol) was added DME (50 mL) and water(10.0 mL) and the resulting mixture was stirred at 100° C. undernitrogen for 1 h. The reaction was cooled to room temperature and mostof the DME was evaporated in vacuo. The residue was partitioned betweenEtOAc and water and the layers were separated. The aqueous phase wasextracted with EtOAc and the combined organic phases were washed withbrine, dried over MgSO₄ and concentrated in vacuo. the residue waspurified by chromatography, eluting with an EtOAc/hexanes gradient;which gave methyl4-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(5.35 g, 12.20 mmol, 94% yield) as a white foam. LCMS (HpH, 2 min),Rt=1.18 min, MH+=439.

Intermediate 314-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid

A solution of methyl4-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 30)(5.23 g, 11.93 mmol) in MeOH (80mL) was treated with aqueous lithium hydroxide (1M, 17.89 mL, 17.89mmol) and the resulting mixture, which became a suspension after 5 minwas stirred. After 24 h, lithium hydroxide (17.89 mL, 17.89 mmol) wasadded and the mixture (which by then was a solution) was stirred at 50°C. for 3 h. The mixture was cooled to room temperature, most of the MeOHwas evaporated in vacuo and the residue treated with acetic acid (2.73mL, 47.7 mmol). After 5 min, a precipitate formed, was isolated byfiltration, washed with water and dried at 45° C. under vacuum for 16 hto give4-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (4.88 g, 11.50 mmol, 96% yield) as a white solid. LCMS (HpH, 2min), Rt=0.70 min, MH+=425.

Intermediate 324-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

A suspension of crushed tert-butyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation see Intermediate 41) (8 g, possibly as the PF6 salt)was treated with 1,4-dioxane (30 mL) followed by hydrogen chloride in1,4-dioxane (4N, 40 mL, 160 mmol). After −30 min most of the dioxane wasremoved and the residue dissolved in warm MeOH (100 mL) at 70° C. for1.5 h. The residual insoluble material which was filtered off anddiscarded. The filtrate was concentrated in vacuo and purified using aSCX column (70 G, pre-conditioned with MeOH.) The cartridge was washedwith MeOH (200 ml) and eluted with ammonia in MeOH (2M, 200 ml). Themethanolic ammonia fractions were combined and concentrated in vacuo togive4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide(3.65 g, 9.25 mmol, 84% yield) as a yellow foam. LCMS (HpH, 2 min),Rt=0.71 min, MH+=395.

Intermediate 33 methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate

6-(((2S,4R)-1-Acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(for a preparation see Intermediate 2)(3.25 g, 8.44 mmol),{6-[(methyloxy)carbonyl]-3-pyridinyl}boronic acid (1.832 g, 10.12 mmol),{6-[(methyloxy)carbonyl]-3-pyridinyl}boronic acid (1.832 g, 10.12 mmol)and triethylamine (2.352 ml, 16.87 mmol) were combined in MeOH (30 ml)and DME (5 ml). The reaction mixture was divided between 2 vials andeach heated at 120° C. for 45 min. The crude reaction mixtures werecombined and concentrated in vacuo to give 4.4 g of crude brown solid.This was purified by chromatography on silica (100 g), eluting with over1500 ml of EtOAc (compound leached off slowly) to give methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(2.94 g, 6.66 mmol, 79% yield) as a brown foamy solid. LCMS (Formate, 2min), Rt=0.81 min, MH+=442.

Intermediate 345-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid

Methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(for a preparation see Intermediate 33)(2.94 g, 6.66 mmol) was dissolvedin anhydrous 1,4-dioxane (20 ml). Water (20.0 ml) was added followed bylithium hydroxide (0.319 g, 13.32 mmol) and reaction mixture stirred atr.t. After 1 h, the 1,4-dioxane was removed in vacuo and aqueous residuewas treated with acetic acid (1.144 ml, 19.98 mmol). The resultingorange precipitate was stirred for 20 min, isolated by filtration,washed with water and dried under vacuum overnight to give5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (2.16 g, 4.55 mmol, 68.3% yield) as a brown solid. LCMS (Formate, 2min), Rt=0.68 min, MH+=428.

Intermediate 354-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid

Pd/C (22.1 mg, 10%, 0.208 mmol, containing ˜50% water) was added to asolution of4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 18)(369 mg, 0.848 mmol) andammonium formate (134 mg, 2.121 mmol) in ethanol (5 ml) and DMF (1 ml)and reaction mixture heated at reflux. After 1.5 h the reaction mixturewas cooled to r.t. The reaction mixture was filtered through Celite™ andfilter cake washed with EtOH. The filtrate was concentrated to give alight brown oil (251 mg). The filter cake was washed with a furtherportion of 1:1 EtOH:DMF (50 ml) and combined with the previous materialand concentrated to give4-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (306 mg, 0.764 mmol, 90% yield) as a yellow solid. LCMS (Formate, 2min), Rt=1.01 min, MH+=401.

Intermediate 36 R-BINAP)ditriflatebis(acetonitrile)palladium(II

R-(+)-BINAP (6.08 g, 9.76 mmol, available from Avocado) was stirred inDCM (626 ml) and dichlorobis(acetonitrile)palladium (II) (2.5 g, 9.64mmol, available from Aldrich) added. The mixture was stirred undernitrogen for 30 min, the suspension had not become a solution and moreDCM (100 ml) was added. The mixture was stirred for a further 30 min andsilver triflate (5.00 g, 19.47 mmol, available from Aldrich) dissolvedin acetonitrile (250 ml) was added. The mixture changed from an orangecloudy suspension to a yellow suspension. The mixture was stirred for 1h, filtered through Celite™ and evaporated to an orange solid. Theresidue was dried under vacuum (at approximately 14 mbar) at roomtemperature over the weekend to give the desired product (10.69 g).

1H NMR (400 MHz, MeCN-d3) δ ppm 2.0 (s, 6H), 6.7 (d, 2H), 6.9 (br m,4H), 7.1 (br t, 2H), 7.2 (t, 2H), 7.5-7.9 (m, 22H)

Intermediate 37 1-Methylethyl{(2S,4R)-1-acetyl-6-[4-({[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}carbonyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate

To a flask charged with4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 48) (82 mg, 0.2 mmol) and HATU(0.091 g, 0.240 mmol) was added DMF (2 mL) and the mixture treated withDIPEA (0.105 mL, 0.600 mmol) at room temperature. After 5 min,1,1-dimethylethyl (2-aminoethyl)carbamate (0.047 mL, 0.300 mmol) wasadded and the mixture stirred at room temperature for 1 h. The reactionmixture was partitioned between EtOAc and water. The phases wereseparated and the aqueous phase was extracted with EtOAc. The combinedorganic layers were washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by chromatographyeluting with a MeOH/DCM gradient to give 1-methylethyl{(2S,4R)-1-acetyl-6-[4-({[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}carbonyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate(95 mg, 0.172 mmol, 86% yield) as a pale brown foam. LCMS (HpH, 2 min),Rt=1.02 min, MH+=553.

Intermediate 38 1-Methylethyl{(2S,4R)-1-acetyl-6-[4-(2-{[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}-2-oxoethyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate

To a flask charged with2-(4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid (for a preparation see Intermediate 50) (85 mg, 0.2 mmol) and HATU(91 mg, 0.240 mmol) was added DMF (2 mL) and the mixture treated withDIPEA (0.105 mL, 0.600 mmol) at room temperature. After 5 min,1,1-dimethylethyl (2-aminoethyl)carbamate (0.047 mL, 0.300 mmol) wasadded, the mixture was stirred at room temperature for 3 h and thenpartitioned between EtOAc and water. The phases were separated and theaqueous phase was extracted with EtOAc. The combined organic phases werewashed with brine, dried over MgSO₄, filtered and concentrated in vacuo.the residue was purified by chromatography, eluting with a MeOH/DCMgradient, to give 1-methylethyl{(2S,4R)-1-acetyl-6-[4-(2-{[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}-2-oxoethyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate(56 mg, 0.099 mmol, 49.4% yield) as a pale yellow foam. LCMS (Formate, 2min), Rt=0.91 min, MH+=455.

Intermediate 39 tert-Butyl(2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate

To a flask charged with4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 4)((70 mg, 0.164 mmol) and HATU(74.9 mg, 0.197 mmol) was added DMF (3 mL) and the resulting mixture wastreated with DIPEA (0.086 mL, 0.492 mmol). After 5 min, tert-butyl(2-aminoethyl)carbamate (52.6 mg, 0.328 mmol) was added and the mixturewas stirred at room temperature for 1 h and then concentrated in vacuo.The residue was partitioned between EtOAc and water and the layers wereseparated. The aqueous phase was extracted with EtOAc (×2) and thecombined organic layers were washed with saturated aqueous NaHCO₃ (×3),then brine, dried over MgSO₄ and concentrated in vacuo to givetert-butyl(2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamateas a brown foam which was further purified by MDAP (HpH). to givetert-butyl(2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate(65 mg, 0.114 mmol, 69.6% yield) as a white solid. LCMS (HpH, 2 min),Rt=1.00 min, MH+=569.

Intermediate 40 tert-butyl(2-(4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate

LCMS (High pH, 2 min), Rt=1.01 min, MH+=544

Intermediate 41 tert-butyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

To a flask charged with4-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 31)(4.88 g, 11.50 mmol) andHATU (5.25 g, 13.80 mmol) was added DMF (2 mL). The resulting mixturewas treated with DIPEA (6.02 mL, 34.5 mmol) then after 5 min withN1,N1-dimethylethane-1,2-diamine (1.317 g, 14.95 mmol). After 0.5 h mostof the DMF was removed in vacuo. The residue was treated with EtOAc andsaturated NaHCO₃. Insoluble solid appeared which was isolated byfiltration and dried under vacuum at 60° C. for 3 h to give tert-butyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamateas white solid (8.5 g), which was used without further purification inthe subsequent reaction. The phases of the filtrate were separated, andthe aqueous extracted with EtOAc (×2). The combined organic phases werewashed with saturated NaHCO₃(×3), dried over MgSO₄ and concentrated invacuo to give a liquid (2.1 g). This liquid was loaded onto a SCXcartridge (50 g, pre-conditioned with MeOH) the cartridge washed withMeOH (200 ml) and eluted with ammonia in MeOH (2M, 200 ml). Themethanolic ammonia fractions were combined and concentrated in vacuo togive a further portion of tert-butyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(0.3 g, 0.6 mmol, 5% yield) as an orange solid. LCMS (Formate, 2 min),Rt=0.94 min, MH+=495.

Intermediate 42 tert-butyl(1-(5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinoyl)azetidin-3-yl)carbamate

LCMS (Formate, 2 min), Rt=0.97 min, MH+=582

Intermediate 43 tert-butyl(1-(5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinoyl)piperidin-4-yl)carbamate

LCMS (Formic, 2 min), Rt=0.97 min, MH+=610

Intermediate 44 tert-butyl4-(5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinoyl)piperazine-1-carboxylate

LCMS (Formate, 2 min), Rt=1.00 min, MH+=596

Intermediate 45 tert-butyl(2-(4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate

LCMS (High pH, 2 min), Rt=1.20 min, MH−=576

Intermediate 46 Methyl4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a solution of isopropyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(5 g, 13.54 mmol) and methyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (3.90 g, 14.89mmol) in DME (50 mL) and water (10.0 mL) were successively addedpalladium tetrakis(triphenylphosphine) (1.565 g, 1.354 mmol) andpotassium carbonate (5.61 g, 40.6 mmol). The resulting mixture wasstirred at 100° C. for 1 h, whereupon it was allowed to cool down toroom temperature and was filtered through Celite™. The filtrated wasconcentrated in vacuo and the residue was partitioned between EtOAc andwater. The aqueous phase was extracted with EtOAc (×3) and the combinedorganic layers were washed with brine, dried over MgSO₄, filtered andconcentrated in vacuo. The crude compound was purified by flashchromatography on a silica gel cartridge (50 g) eluting with EtOAc incyclohexane (5-60%). The appropriate fractions were combined andconcentrated under reduced pressure to give methyl4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(4.64 g, 81%) as a white gum. LCMS (Formate, 2 min), Rt=1.09 min,MH+=425.

Intermediate 47

Lithium4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

To a solution of methyl4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 46)(1.63 g, 3.84 mmol) in MeOH (20mL) was added lithium hydroxide (4.61 mL, 4.61 mmol). The resultingmixture was stirred at 40° C. for 6 h, whereupon it was concentratedunder reduced pressure to give lithium4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(1.65 g, 100%) which was not purified but directly used in thesubsequent step. LCMS (Formate, 2 min), Rt=0.87 min, MH+=411.

Intermediate 484-((2S,4R)-1-Acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid

Lithium4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(1.05 g, 2.52 mmol) (for a preparation see Intermediate 47) waspartitioned between EtOAc and hydrochloric acid (2M). The phases wereseparated and the aqueous phase was extracted with EtOAc (×3). Thecombined organic layers were washed with brine, dried over MgSO₄,filtered and concentrated in vacuo to give4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (898 mg, 87%) as a white solid. LCMS (Formate, 2 min), Rt=0.87 min,MH+=411.

Intermediate 49 Ethyl2-(4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)acetate

To a flask was charged with ethyl (4-bromophenyl)acetate (0.174 mL,1.000 mmol), 1-methylethyl[(2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydro-4-quinolinyl]carbamate(416 mg, 1 mmol), potassium carbonate (415 mg, 3.00 mmol) andPdCl₂(dppf) (73.2 mg, 0.100 mmol) was added 1,4-dioxane (6 mL) and water(2.0 mL) and the flask flushed with nitrogen. The resulting mixture wasstirred under microwave irradiation at 120° C. for 30 min then cooled toroom temperature. The bulk of dioxane was removed in vacuo and theresidue partitioned between EtOAc and water. The layers were separatedand the aqueous phase was extracted with EtOAc. The combined organiclayers were washed with brine, dried over MgSO₄ and concentrated invacuo. The residue was purified by chromatography [(25 g column,MeOH/DCM)] to give ethyl2-(4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)acetate(270 mg, 59.7% yield). This compound was used in the next step withoutfurther purification. LCMS (HpH, 2 min), Rt=1.16, MH+=453.

Intermediate 502-(4-((2S,4R)-1-Acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid

To a solution of ethyl{4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]phenyl}acetate(for a preparation see Intermediate 49)(270 mg, 0.597 mmol) in MeOH (6mL) and water (2.0 mL) was added aqueous sodium hydroxide (2N, 0.597 mL,1.193 mmol) at room temperature and the resulting mixture was stirredfor 6 h. Aqueous sodium hydroxide (2N, 0.5 mL) was added and the mixturewas left standing overnight. The bulk of MeOH was removed in vacuo andthe resulting residue was partitioned between water and Et₂O and thelayers were separated. The aqueous layer was acidified with hydrochloricacid (2N, 2 mL) and extracted twice with EtOAc. The combined organicphases were dried over MgSO₄ and concentrated in vacuo to give{4-[(2S,4R)-1-acetyl-2-methyl-4-({[(1-methylethyl)oxy]carbonyl}amino)-1,2,3,4-tetrahydro-6-quinolinyl]phenyl}aceticacid (200 mg, 0.471 mmol, 79% yield) as a brown foam. This compound wasused in the next step without further purification. LCMS (HpH, 2 min),Rt=0.65, MH+=425.

Intermediate 515-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)pyrazine-2-carbonitrile

1-((2S,4R)-4-amino-6-bromo-2-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 1) (50 mg, 0.177 mmol),5-chloropyrazine-2-carbonitrile (37.0 mg, 0.265 mmol) and DIPEA (0.123mL, 0.706 mmol) were combined in NMP (1.5 mL) and heated in a sealedflask under microwave irradiation to 200° C. for 1 h. The mixture waspartitioned between water (150 mL) and EtOAc (4×100 mL). The organiclayers were combined, washed with brine (2×100 mL), dried by passingthrough a hydrophobic frit, and concentrated in vacuo. The resultingresidue was purified by chromatography (25 g column, EtOAc/cyclohexanegradient) to give5-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)pyrazine-2-carbonitrile(61 mg, 0.158 mmol, 89% yield) as a brown gum.

LCMS (Formate, 2 min), Rt=0.94 mins, MH+=386/388.

Intermediate 52 Methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate

5-(((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)pyrazine-2-carbonitrile(for a preparation, see Intermediate 51) (390 mg, 1.010 mmol),(6-(methoxycarbonyl)pyridin-3-yl)boronic acid (219 mg, 1.21 mmol),PdCl₂(dppf) (111 mg, 0.151 mmol) and triethylamine (0.281 ml, 2.02 mmol)were combined in a mixture of dry MeOH (6 mL) and DME (1 mL). Themixture was heated under microwave irradiation to 120° C. for 45 mins.The mixture was then concentrated in vacuo and the residue purified bychromatography (100 g column, EtOAc/cyclohexane gradient) to give methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(389 mg, 0.879 mmol, 87% yield) as a pink solid.

LCMS: (Formate, 2 min), Rt=0.84 mins, MH+=443.

Intermediate 535-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid

Lithium hydroxide monohydrate (83 mg, 1.97 mmol) in water (5 mL) wasadded to methyl5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(for a preparation, see Intermediate 52) (435 mg, 0.983 mmol) in1,4-dioxane (5 mL) and the reaction mixture stirred at room temperaturefor 70 mins. Acetic acid (0.169 mL, 2.95 mmol) was added and theresulting mixture stirred for 10 mins. The mixture was filtered, thesolid collected was washed with water, then dried in a vacuum oven togive5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (181 mg, 0.422 mmol, 43% yield) as a brown solid. LCMS: (Formate, 2min) Rt=0.66 mins, MH+=429.

Intermediate 54 Methyl5-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate

(2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinamine(for a preparation, see Intermediate 1) (600 mg, 1.88 mmol), methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinate (593 mg, 2.25mmol), PdCl₂(dppf) (206 mg, 0.282 mmol) and triethylamine (0.785 mL,5.63 mmol) were combined in DME (3 mL) and MeOH (12 mL) and the mixtureheated under microwave irradiation to 110° C. for 45 mins.

In a separate vial,(2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydro-4-quinolinamine(for a preparation, see Intermediate 1) (50 mg, 0.156 mmol), methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinate (49.4 mg,0.188 mmol), PdCl₂(dppf) (17.17 mg, 0.023 mmol) and triethylamine (0.065mL, 0.469 mmol) were combined in DME (0.5 mL) and MeOH (1 mL) and themixture heated under microwave irradiation to 110° C. for 45 mins. Thetwo crude reaction mixtures were combined and concentrated in vacuo. Theresidue was purified by chromatography (100 g column, EtOAc/cyclohexanefollowed by 2M methanolic ammonia/DCM gradient). The appropriatefractions were combined, concentrated in vacuo and dried in a vacuumoven to give a residue. This residue was partitioned between sat. aq.sodium bicarbonate (100 mL) and DCM (3×100 mL). The organic layers werecombined, dried by passing through a hydrophobic frit and concentratedin vacuo to give methyl5-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(555 mg, 1.64 mmol, 87% yield).

LCMS: (Formate, 2 min), Rt=0.53 mins, MH+=340.

Intermediate 555-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid

To methyl5-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinate(for a preparation, see Intermediate 54) (550 mg, 1.62 mmol) in MeOH (5mL) was added lithium hydroxide monohydrate (78 mg, 3.24 mmol) in water(5 mL) and the mixture stirred at room temperature for 90 mins. Themixture was concentrated in vacuo before DCM was added. The mixture wasfiltered, dried by passing through a hydrophobic frit, and concentratedin vacuo to give5-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (517 mg, 1.59 mmol, 98% yield).

LCMS: (Formate, 2 min), Rt=0.38 mins, MH+=326.

Intermediate 561-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

5-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (for a preparation, see Intermediate 55) (510 mg, 1.567 mmol),morpholine (0.149 mL, 1.72 mmol), HATU (656 mg, 1.724 mmol) and DIPEA(0.821 mL, 4.70 mmol) were combined in DMF (5 mL) and the mixturestirred at room temperature overnight. Morpholine (0.149 mL, 1.72 mmol)was added and the reaction stirred at room temperature for a further 3h. The mixture was concentrated in vacuo before being dissolved in DCMand loaded onto an aminopropyl SPE column (50 g). The column was elutedwith 10% MeOH in DCM and the appropriate fractions were combined andconcentrated in vacuo to give a residue. This residue was purified bychromatography (50 g column, 2M methanolic ammonia/DCM gradient) to give1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(470 mg, 1.19 mmol, 76% yield) as a beige solid. LCMS: (Formate, 2 min),Rt=0.48 mins, MH+=395.

Intermediate 57 Ethyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

Sodium tert-butoxide (135 mg, 1.41 mmol)) was added to a suspension ofethyl4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 16) (450 mg, 1.28 mmol),2-bromopyridine (403 mg, 2.55 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (40 mg,0.102 mmol) and tris(dibenzyldieneacetone)dipalladium(0) (47 mg, 0.051mmol) in anhydrous toluene (4.5 mL). The mixture was heated undermicrowave irradiation to 120° C. for 1 h in a sealed vial. The mixturewas partitioned between EtOAc and water, the aqueous layer was extractedtwice with EtOAc, the combined organic phases were dried by passingthrough a hydrophobic frit and concentrated in vacuo to give a residue.The residue was purified by chromatography (10 g column,EtOAc/cyclohexane gradient followed by MeOH/EtOAc gradient) to giveethyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(216 mg, 0.503 mmol, 39% yield) as a brown gum. LCMS: (Formate, 2 min),Rt=0.84 min, MH+430.

Intermediate 58 Lithium4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

Ethyl4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 57) (216 mg, 0.503 mmol) in EtOH (1mL) and water (0.5 mL) was treated with lithium hydroxide monohydrate(22 mg, 0.524 mmol) and the mixture stirred at r.t. over the weekend. Afurther portion of lithium hydroxide monohydrate (4 mg, 0.095 mmol) wasadded to the reaction and the mixture stirred for 1 h. The mixture wasconcentrated to dryness to give the crude product lithium4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(160 mg) which was used directly in the next step without furtherpurification.

LCMS: (Formate, 2 min), Rt=0.62 min, MH+402.

Intermediate 59 Ethyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

Sodium tert-butoxide (135 mg, 1.41 mmol)) was added to a suspension ofethyl4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 16) (450 mg, 1.28 mmol),2-bromo-5-methylpyridine (439 mg, 2.55 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (40 mg,0.102 mmol) and tris(dibenzyldieneacetone)dipalladium(0) (47 mg, 0.051mmol) in anhydrous toluene (4.5 mL). The mixture was heated undermicrowave irradiation to 120° C. for 1 h in a sealed vial. The mixturewas partitioned between EtOAc and water, the aqueous layer was extractedtwice with EtOAc, the combined organic phases were dried by passingthrough a hydrophobic frit and concentrated in vacuo to give a residue.The residue was purified by chromatography (10 g column,EtOAc/cyclohexane gradient followed by MeOH/EtOAc gradient) to give aresidue. This residue was dissolved in a mixture of EtOAc (0.6 mL) andcyclohexane (0.4 mL). The solution was treated with cyclohexane (3 mL)to give an emulsion, which was allowed to stand at room temperature overa weekend. The solution was decanted from the precipitated dark brownoil and concentrated to dryness under a stream of nitrogen to give anoil. This oil was triturated with cyclohexane (2×3 mL), the residual gumisolated by decanting off the cyclohexane, dissolved in EtOAc andconcentrated to dryness under a stream of nitrogen to give crude ethyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoateas a pale brown foam (240 mg) which was used directly in the next stepwithout further purification.

LCMS: (Formate, 2 min), Rt=0.88 min, MH+444.

Intermediate 60

Lithium4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

Crude ethyl4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(Intermediate 59) (238 mg) in EtOH (2 mL) and water (1 mL) was treatedwith lithium hydroxide monohydrate (26 mg, 0.620 mmol). The solution wasstirred at ambient temperature for 5 h. A further portion of lithiumhydroxide monohydrate (4 mg, 0.095 mmol) was added and the mixture leftat ambient temperature overnight. The mixture was concentrated todryness to give crude lithium4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoateas a brown foam (240 mg) which was used directly in the next stepwithout further purification. LCMS: (Formate, 2 min), Rt=0.65 min,MH+416.

Intermediate 61 Tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

Tert-butyl((2S,4R)-1-acetyl-6-bromo-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation, see Intermediate 29) (2.0 g, 5.22 mmol),bis(pinacolato)diboron (2.92 g, 11.48 mmol), PdCl₂(dppf) (0.191 g, 0.261mmol) and potassium acetate (1.54 g, 15.65 mmol) were combined in1,4-dioxane (25 mL) and degassed. The mixture was then heated under anitrogen atmosphere for 75 mins at 105° C. The mixture was concentratedin vacuo and the residue purified by chromatography (100 g column,EtOAc/cyclohexane) to give tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(2.36 g, 4.94 mmol, 95% yield).

LCMS: (Formate, 2 min), Rt=1.21 mins, MH+=431.

Intermediate 62 Methyl6-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate

Tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation see Intermediate 61) (2.2 g, 5.11 mmol), methyl6-bromonicotinate (1.325 g, 6.13 mmol), Pd(PPh₃)₄ (0.591 g, 0.511 mmol)and potassium carbonate (2.120 g, 15.34 mmol) were combined in methanol(10 mL) and DME (3 mL) and heated under microwave irradiation to 100° C.for 70 mins.

In a separate vial, Tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation see Intermediate 61) (2.2 g, 5.11 mmol), methyl6-bromonicotinate (1.325 g, 6.13 mmol), Pd(PPh₃)₄ (0.591 g, 0.511 mmol)and potassium carbonate (2.120 g, 15.34 mmol) were combined in methanol(10 mL) and DME (3 mL) and heated under microwave irradiation to 100° C.for 70 mins. The two reaction mixtures were combined and partitionedbetween water (150 mL) and DCM (3×150 mL). The aqueous layer wasacidified to pH 3 via the addition of aq. HCl and extracted with DCM(3×100 mL). The combined organic layers were dried by passing through ahydrophobic frit, and concentrated in vacuo to give crude methyl6-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(2.2 g) which was used directly in the next step without furtherpurification. LCMS: (Formate, 2 min), Rt=1.06 min, MH+440.

Intermediate 636-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid

To a stirring solution of crude methyl6-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinate(Intermediate 62) (2.2 g, 5.01 mmol) in MeOHl (12 mL), was added lithiumhydroxide monohydrate (0.144 g, 6.01 mmol) in water (12 mL) and themixture warmed to 50° C. for 70 mins. MeOH (10 mL) was added and themixture stirred at 50° C. for a further 50 mins. The mixture was allowedto cool to room temperature before being partitioned between saturatedaqueous sodium bicarbonate (150 mL) and DCM (3×150 mL). The aqueouslayer was then treated with 2M HCl until acidic (˜pH 3) before beingextracted with DCM (3×150 ml). The organics were combined, dried bypassing through a hydrophobic frit and concentrated in vacuo to give6-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (2.08 g, 4.89 mmol). LCMS: (Formate, 2 min), Rt=0.86 mins, MH+426.

Intermediate 64 Tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

6-((2S,4R)-1-acetyl-4-((tert-butoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (for a preparation, see Intermediate 63) (2.05 g, 4.82 mmol),morpholine (2.084 mL, 24.09 mmol), HATU (2.015 g, 5.30 mmol) and DIPEA(2.52 mL, 14.45 mmol) were combined in DMF (25 mL) and the reactionstirred at room temperature for 90 mins. The mixture was concentrated invacuo before being dissolved in DCM and loaded onto an aminopropyl SPEcolumn (70 g). The column was eluted with 10% MeOH in DCM, and theappropriate fractions combined and concentrated to dryness in cavuo. Theresidue was purified by chromatography (100 g column, EtOAc/cyclohexanegradient) to give tert-butyl((2S,4R)-1-acetyl-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(2.05 g, 4.14 mmol, 86% yield).

LCMS: (Formate, 2 min), Rt=0.86 mins, MH+495.

Intermediate 651-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride

4M hydrochloric acid in 1,4-dioxane (10.11 ml, 40.4 mmol) was added totert-butyl((2S,4R)-1-acetyl-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(for a preparation, see Intermediate 64) (2 g, 4.04 mmol) at roomtemperature, followed by MeOH (10 mL) and DCM (10 mL) and the mixturestirred at room temperature for 2 mins. The mixture was concentrated invacuo to give1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (1.60 g, 3.71 mmol, 92% yield).

LCMS: (Formate, 2 min), Rt=0.50 mins, MH+395.

Intermediate 66 Ethyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

Ethyl4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation, see Intermediate 16) (300 mg, 0.851 mmol) and6-chloronicotinonitrile (236 mg, 1.702 mmol) in anhydrous NMP (5 mL)were treated with DIPEA (0.445 mL, 2.55 mmol). The reaction vial wassealed and heated under microwave irradiation to 200° C. for 2 h. Thecooled solution was diluted with water to −20 mL, stirred and theresulting sticky solid isolated by decantation. The solid was dissolvedin EtOAc, the solution washed with brine and dried by passing through ahydrophobic frit. The solution was concentrated in vacuo and the residuepurified by MDAP (Formate) to give crude ethyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(120 mg) as a beige gum which was used directly in the next step withoutfurther purification. LCMS: (Formate, 2 min), Rt=1.12 min, MH+455.

Intermediate 67 Lithium4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate

A solution of crude ethyl4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(Intermediate 66) (120 mg) in EtOH (2 mL) was treated with lithiumhydroxide monohydrate (13 mg, 0.303 mmol) and water (1 mL). The mixturewas stirred at room temperature for ˜24 h. The solution was concentratedto dryness to give crude lithium4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoatecrudeas a crunchy yellow foam (110 mg) which was used directly in the nextstep without further purification.

LCMS: (Formate, 2 min), Rt=0.87 min, MH+427.

Example 13-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)propanamide

To a solution of3-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)propanoicacid (for a preparation See Intermediate 10)(58 mg, 0.128 mmol) andN1,N1-dimethylethane-1,2-diamine (22.50 mg, 0.255 mmol) in DMF (3 mL)were added HATU (72.8 mg, 0.191 mmol) and DIPEA (0.067 mL, 0.383 mmol).The resulting mixture was stirred for 30 min, then reaction mixturepartitioned between water and EtOAc. The aqueous phase was extractedwith EtOAc (×3), the combined organic layers were washed with brine,dried over MgSO₄, filtered and concentrated in vacuo. The crude residuewas dissolved in 1:1 MeOH/DMSO and was purified by MDAP (formate). Theappropriate fractions were combined and concentrated in vacuo to give3-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)propanamide(34.6 mg, 48%) as a viscous colourless oil. LCMS (Formate, 2 min),Rt=0.71 min, MH+=525.

Example 23-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-hydroxyethyl)propanamide

To a solution of3-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)propanoicacid (for a preparation see Intermediate 10)(44 mg, 0.097 mmol) andethanolamine (0.012 mL, 0.194 mmol) in DMF (3 mL) were added HATU (55.2mg, 0.145 mmol) and DIPEA (0.051 mL, 0.290 mmol). The resulting mixturewas stirred for 40 min, then partitioned between water and EtOAc. Theaqueous phase was extracted with EtOAc (×3), the combined organic layerswere washed with brine, dried over MgSO₄, filtered and concentrated invacuo. The crude residue was dissolved in 1:1 MeOH/DMSO and was purifiedby MDAP (formate). The appropriate fractions were combined andconcentrated in vacuo to give3-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-hydroxyethyl)propanamide(22.8 mg, 47%) as a, viscous yellow oil. LCMS (Formate, 2 min), Rt=0.80min, MH+=498.

Example 32-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-hydroxyethyl)acetamide

To a solution of2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid, lithium salt (Intermediate 9)(166 mg, 0.377 mmol) and ethanolamine(0.034 mL, 0.565 mmol) in DMF (3 mL) were added HATU (287 mg, 0.754mmol) and DIPEA (0.197 mL, 1.131 mmol). The resulting mixture wasstirred at room temperature for 1 h, the reaction mixture was dilutedwith EtOAc and water. The layers were separated and the aqueous phasewas extracted with EtOAc (×3). The combined organic layers were washedwith brine, dried over MgSO₄, filtered and concentrated in vacuo. Thecrude residue was dissolved in 1:1 MeOH/DMSO and was purified by MDAP(formate). The appropriate fractions were combined and concentrated invacuo to give2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-hydroxyethyl)acetamide(21 mg, 11%) as a colourless oil. LCMS (Formate, 2 min), Rt=0.76 min,MH+=484.

Example 42-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(1,3-dihydroxypropan-2-yl)acetamide

To a solution of2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid, lithium salt (for a preparation see Intermediate 9)(169 mg, 0.384mmol) and 2-aminopropane-1,3-diol (69.9 mg, 0.767 mmol) in DMF (2 ml)were added HATU (190 mg, 0.499 mmol) and DIPEA (0.201 mL, 1.151 mmol).The resulting mixture was stirred at room temperature for 2 h. Thereaction mixture was diluted with EtOAc and water. The layers wereseparated and the aqueous phase was extracted with EtOAc (×3). Thecombined organic layers were washed with brine, dried over MgSO₄,filtered and concentrated in vacuo. The crude residue was dissolved in1:1 MeOH/DMSO and was purified by MDAP (formate). The appropriatefractions were combined and concentrated in vacuo to give2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(1,3-dihydroxypropan-2-yl)acetamide(83 mg, 42%) as a colourless oil. LCMS (Formate, 2 min), Rt=0.73 min,MH+=514.

Example 52-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)acetamide

To a solution of2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)aceticacid, lithium salt (for a preparation see Intermediate 9)(125 mg, 0.284mmol) and N1,N1-dimethylethane-1,2-diamine (32.5 mg, 0.369 mmol) in DMF(3 mL) were added HATU (162 mg, 0.426 mmol) and DIPEA (0.149 mL, 0.851mmol). The resulting mixture was stirred at room temperature for 2 h,the reaction mixture was diluted with EtOAc and water. The layers wereseparated and the aqueous phase was extracted with EtOAc (×3). Thecombined organic layers were washed with brine, dried over MgSO₄,filtered and concentrated in vacuo. The crude residue was dissolved in1:1 MeOH/DMSO and was purified by MDAP (formate). The appropriatefractions were combined and concentrated in vacuo to give2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)acetamide(59 mg, 41%) as a colourless oil. LCMS (Formate, 2 min), Rt=0.65 min,MH+=511.

Example 6 Isopropyl((2S,4R)-1-acetyl-6-(4-((2-aminoethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamatehydrochloride

A solution of 1-methylethyl{(2S,4R)-1-acetyl-6-[4-({[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}carbonyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate(for a preparation see Intermediate 37)(90 mg, 0.163 mmol) in dioxane (2ml) was treated with hydrogen chloride in 1,4-dioxane (4N, 2 mL, 8.00mmol) and the resulting mixture stirred at room temperature for 5 h,then concentrated in vacuo. The resulting residue was triturated withEt₂O and isolated by filtration to give isopropyl((2S,4R)-1-acetyl-6-(4-((2-aminoethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamatehydrochloride (68 mg, 0.139 mmol, 85% yield) as a pale yellow solid.LCMS (HpH, 2 min), Rt=0.78 min.

Example 7 Isopropyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

To a solution of4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 48) (67 mg, 0.163 mmol) andN1,N1-dimethylethane-1,2-diamine (28.8 mg, 0.326 mmol) in DMF (2 mL)were added HATU (93 mg, 0.245 mmol) and DIPEA (0.086 mL, 0.490 mmol).The resulting mixture was stirred at room temperature for 1 h, thenhydrolyzed by adding water and EtOAc. The layers were separated and theaqueous phase was extracted with EtOAc (×3). The combined organic layerswere washed with brine, dried over MgSO₄, filtered and concentrated invacuo. The residue was dissolved in 1:1 MeOH/DMSO and was purified byMDAP (formate). The appropriate fractions were combined and concentratedunder reduced pressure to give isopropyl((2S,4R)-1-acetyl-6-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(35 mg, 41%) as a viscous colourless oil. LCMS (Formate, 2 min), Rt=0.65min, MH+=481.

Example 8 Isopropyl((2S,4R)-1-acetyl-6-(4-((2-hydroxyethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate

To a solution of4-((2S,4R)-1-acetyl-4-((isopropoxycarbonyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 48) (116 mg, 0.283 mmol) andethanolamine (0.034 mL, 0.565 mmol) in DMF (3 mL) were added HATU (161mg, 0.424 mmol) and DIPEA (0.148 mL, 0.848 mmol). The resulting mixturewas stirred at room temperature for 1 h, then water and EtOAc wereadded. The layers were separated and the aqueous phase was extractedwith EtOAc (×3). The combined organic layers were washed with brine,dried over MgSO₄, filtered and concentrated in vacuo. The residue wasdissolved in 1:1 MeOH/DMSO and was purified by MDAP (formate). Theappropriate fractions were combined and concentrated under reducedpressure to give isopropyl((2S,4R)-1-acetyl-6-(4-((2-hydroxyethyl)carbamoyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamate(102 mg, 80%) as a white solid. LCMS (Formate, 2 min), Rt=0.76 min,MH+=454.

Example 9 Isopropyl((2S,4R)-1-acetyl-6-(4-(2-((2-aminoethyl)amino)-2-oxoethyl)phenyl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)carbamatehydrochloride

A solution of 1-methylethyl{(2S,4R)-1-acetyl-6-[4-(2-{[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)ethyl]amino}-2-oxoethyl)phenyl]-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl}carbamate(for a preparation see Intermediate 38) (60 mg, 0.106 mmol) in1,4-dioxane (2 ml) was treated with hydrogen chloride in 1,4-dioxane(4N, 2 mL, 8.00 mmol) and the resulting mixture stirred at roomtemperature for 5 h then concentrated in vacuo. The residue wastriturated with Et₂O and the solid isolated by filtration to give1-methylethyl[(2S,4R)-1-acetyl-6-(4-{2-[(2-aminoethyl)amino]-2-oxoethyl}phenyl)-2-methyl-1,2,3,4-tetrahydro-4-quinolinyl]carbamatehydrochloride (28 mg, 0.056 mmol, 52.6% yield) as a pale yellow solid.LCMS (HpH, 2 min), Rt=0.79 min, MH+=467.

Example 106-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide

To a flask charged with6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (for a preparation see Intermediate 7)(90 mg, 0.211 mmol) and HATU(96 mg, 0.253 mmol) was added DMF (3 mL) and the resulting mixture wastreated with DIPEA (0.110 mL, 0.632 mmol). After stirring for 5 min, themixture was treated with N1,N1-dimethylethane-1,2-diamine (37.1 mg,0.421 mmol). After stirring for 3 min, the solution was partitionedbetween EtOAc and saturated aqueous NaHCO₃ solution and the layers wereseparated. The aqueous phase was extracted with EtOAc (×2) and thecombined organic layers were washed with saturated aqueous NaHCO₃ (×3),then dried over MgSO₄ and concentrated in vacuo. The residue waspurified by MDAP (HpH). The appropriate fractions were collected andconcentrated in vacuo. The residue was subsequently dissolved in MeOHand dried (hydrophobic frit). The solvent was removed and the residuetriturated in Et₂O to give6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide(36 mg, 0.072 mmol, 34.4% yield) as a white solid.

LCMS (HpH, 2 min), Rt=0.73 min, MH+=471.

Example 116-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)nicotinamide

To a flask charged with6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (for a preparation see Intermediate 7) (90 mg, 0.211 mmol) and HATU(96 mg, 0.253 mmol) was added DMF (3 mL) and the resulting mixture wastreated with DIPEA (0.110 mL, 0.632 mmol). After stirring for 5 min, themixture was treated with 2-aminopropane-1,3-diol (38.4 mg, 0.421 mmol).After stirring for 3 min, the solution was partitioned between EtOAc andsaturated aqueous NaHCO₃ solution and the layers were separated. Theaqueous phase was extracted with EtOAc (×2) and the combined organiclayers were washed with saturated aqueous NaHCO₃ solution (×3), thendried over MgSO₄ and concentrated in vacuo. The residue was purified byMDAP (HpH). The appropriate fractions were collected and concentrated invacuo. The residue was subsequently dissolved in MeOH and dried(hydrophobic frit). The solvent was removed and the residue trituratedin Et₂O to give6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)nicotinamide(26 mg, 0.052 mmol, 24.67% yield) as a white solid.

LCMS (HpH, 2 min), Rt=0.81 min, MH+=498.

Example 126-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)nicotinamide

To a flask charged with6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (for a preparation see Intermediate 7)(90 mg, 0.211 mmol) and HATU(96 mg, 0.253 mmol) was added DMF (3 mL) and the resulting mixture wastreated with DIPEA (0.110 mL, 0.632 mmol). After stirring for 5 min, themixture was treated with 2-aminoethanol (0.013 mL, 0.211 mmol). Afterstirring for 3 min, the solution was partitioned between EtOAc andsaturated aqueous NaHCO₃ solution and the layers were separated. Theaqueous phase was extracted with EtOAc (×2) and the combined organiclayers were washed with saturated aqueous NaHCO₃ solution (×3), thendried over MgSO₄ and concentrated in vacuo. The residue was purified byMDAP (HpH). The appropriate fractions were collected and concentrated invacuo. The residue was subsequently dissolved in MeOH and dried(hydrophobic frit). The solvent was removed and the residue trituratedin Et₂O to give6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)nicotinamide(37 mg, 0.079 mmol, 37.3% yield) as a white solid. LC/MS (HpH, 2 min),Rt=0.70 min, MH+=501.

Example 134-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((S)-2,3-dihydroxypropyl)benzamide

A solution of4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 4) (90 mg, 0.211 mmol) in DMF(2 mL) was treated with EDC (48.5 mg, 0.253 mmol), HOBT hydrochloride(38.8 mg, 0.253 mmol) and N-ethylmorpholine (0.080 mL, 0.633 mmol) andafter 3 min with (S)-3-aminopropane-1,2-diol (38.5 mg, 0.422 mmol). Theresulting mixture was stirred at room temperature for 5 h then EDC (48.5mg, 0.253 mmol), N-ethyl morpholine (0.080 mL, 0.633 mmol), HOBT (38.8mg, 0.253 mmol) and further (S)-3-aminopropane-1,2-diol (38.5 mg, 0.422mmol) were added and the corresponding mixture was stirred for 16 h thenconcentrated in vacuo. The residue was partitioned between water andEtOAc and the layers were separated. The aqueous phase was extractedwith EtOAc (×2) and the combined organic layers were washed withsaturated aqueous NaHCO₃ (×3), then brine, dried over MgSO₄ andconcentrated in vacuo. The residue was purified by MDAP to give4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((S)-2,3-dihydroxypropyl)benzamide(30 mg, 29%) as a white solid. LCMS (HpH, 2 min), Rt=0.75 min, MH+=500.

Example 144-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)benzamide

A solution of4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 4) (90 mg, 0.211 mmol) in DMF(2 mL) was treated with EDC (48.5 mg, 0.253 mmol), HOBT hydrochloride(38.8 mg, 0.253 mmol) and N-ethylmorpholine (0.080 mL, 0.633 mmol) andafter 3 min with 2-aminoethanol (0.025 mL, 0.422 mmol)). The resultingmixture was stirred at room temperature for 5 h then EDC (48.5 mg, 0.253mmol), N-ethyl morpholine (0.080 mL, 0.633 mmol), HOBT (38.8 mg, 0.253mmol) and further 2-aminoethanol (0.025 mL, 0.422 mmol) were added andthe corresponding mixture was stirred for 16 h then concentrated invacuo. The residue was partitioned between water and EtOAc and thelayers were separated. The aqueous phase was extracted with EtOAc (×2)and the combined organic layers were washed with saturated aqueousNaHCO₃ (×3), then brine, dried over MgSO₄ and concentrated in vacuo. Theresidue was purified by MDAP to give4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)benzamide(8 mg, 8%) as a white solid. LCMS (HpH, 2 min), Rt=0.78 min, MH+=470.

Example 154-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide

To a flask charged with4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 4) (90 mg, 0.211 mmol) and HATU(96 mg, 0.253 mmol) was added DMF (3 mL) and the resulting mixture wastreated with DIPEA (0.111 mL, 0.633 mmol). After 5 min, the mixture wastreated with 2-aminopropane-1,3-diol (38.5 mg, 0.422 mmol). The reactionmixture was stirred for 1 h, whereupon the bulk of solvent was removedand the residue was partitioned between EtOAc and saturated aqueousNaHCO₃. The phases were separated and the aqueous phase was extractedwith EtOAc (×2). The combined organic layers were washed with saturatedaqueous NaHCO₃ (×3), then brine, dried over MgSO₄ and concentrated invacuo. The residue was purified by MDAP (HpH). The appropriate fractionswere collected and concentrated in vacuo. The residue was dissolved inMeOH and dried (hydrophobic frit). The solvent was then removed in vacuoand the residue triturated with Et₂O to give4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide(32 mg, 0.064 mmol, 30% yield) as a white solid. LCMS (HpH, 2 min),Rt=0.75 min, MH+=500.

Example 164-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamidedihydrochloride

To a flask charged with4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 4)(100 mg, 0.234 mmol) and HATU(107 mg, 0.281 mmol) was added DMF (2 mL) and the resulting mixture wastreated with DIPEA (0.123 mL, 0.703 mmol). After 5 min,N1,N1-dimethylethane-1,2-diamine (41.3 mg, 0.469 mmol) was added and themixture was stirred for 30 min. The bulk of DMF was removed and theresidue was partitioned between EtOAc and saturated aqueous NaHCO₃. Thephases were separated and the organic layer was washed with saturatedaqueous NaHCO₃, dried over MgSO₄ and concentrated in vacuo. The residuewas purified by MDAP (HpH). The appropriate fractions were collected andconcentrated in vacuo. The residue was co-evaporated with toluene,dissolved in 1,4-dioxane (4 mL) and subsequently treated with hydrogenchloride in 1,4-dioxane (4N, 1 ml). The resulting mixture was stirredfor 5 min at room temperature and concentrated in vacuo. The obtainedresidue was triturated with Et₂O then dried under vacuum to give4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide,di hydrochloride (38 mg, 0.067 mmol, 28.5% yield) as a very pale yellowsolid. LCMS (High pH, 2 min), Rt=0.87 min, MH+=497.

Example 174-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-aminoethyl)benzamidehydrochloride

A suspension of tert-butyl(2-(4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate(for a preparation see Intermediate 39)(65 mg, 0.114 mmol) in1,4-dioxane (1 mL) at room temperature was treated with hydrogenchloride in 1,4-dioxane (4N, 2 mL, 8.00 mmol) and the resultingsuspension was stirred at this temperature for 1 h then concentrated invacuo. The precipitate formed was triturated with Et₂O isolated byfiltration and dried under vacuum to give4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-aminoethyl)benzamide,hydrochloride (60 mg, 0.119 mmol, 104% yield) as a white solid. LCMS(HpH, 2 min), Rt=0.75 min, MH+=469.

Example 184-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-((S)-2,3-dihydroxypropyl)benzamide

A solution of4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 23) (70 mg, 0.174 mmol) in DMF(2 mL) was treated with EDC (40.1 mg, 0.209 mmol), HOBT (32.0 mg, 0.209mmol) and N-ethylmorpholine (0.066 mL, 0.523 mmol) and after 3 min with(S)-3-aminopropane-1,2-diol (15.89 mg, 0.174 mmol). The resultingmixture was stirred at room temperature for 5 h then further EDC (40.1mg, 0.209 mmol), N-ethylmorpholine (0.066 mL, 0.523 mmol), HOBT (32.0mg, 0.209 mmol) and (S)-3-aminopropane-1,2-diol (15.89 mg, 0.174 mmol)were added and the mixture stirred for 16 h before concentrating invacuo. The residue was partitioned between water and EtOAc and thelayers were separated. The aqueous phase was extracted with EtOAc andthe combined organic layers were washed with saturated aqueous NaHCO₃(×3) then brine, dried over MgSO₄ and concentrated in vacuo. Triturationof the residue with Et₂O gave4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N—((S)-2,3-dihydroxypropyl)benzamide(36 mg, 0.076 mmol, 43.5% yield) as a pale yellow solid.

LCMS (HpH, 2 min), Rt=0.76 min, MH+=475

The following analogues were prepared by using the above method with theappropriate amine:—

Example 194-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)benzamide

LC/MS (HpH, 2 min), Rt=0.78 min, MH+=445

Example 204-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

LC/MS (HpH, 2 min), Rt=0.89 min, MH+=472

Example 214-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide

To a flask charged with4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 23) (70 mg, 0.174 mmol) andHATU (80 mg, 0.209 mmol) was added DMF (3 mL) and the resulting solutionwas treated with DIPEA (0.091 mL, 0.523 mmol) and after 5 min by2-aminopropane-1,3-diol (31.8 mg, 0.349 mmol). The reaction wasconcentrated in vacuo, residue partitioned between water and EtOAc. thephases were separated and the aqueous extracted with EtOAc (×2). Thecombined organic layers were washed with saturated NaHCO₃ (3×) thenbrine, dried over MgSO₄ and concentrated in vacuo. The residue waspurified by MDAP (HpH) and the residue triturated with Et₂O to give4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide(17 mg, 0.036 mmol, 20.55% yield) as a very pale yellow solid. LCMS(HpH, 2 min), Rt=0.74 min, MH+=475.

The following analogue was prepared by using the above method with theappropriate amine:—

Example 224-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-aminoethyl)benzamide

A suspension of tert-butyl(2-(4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzamido)ethyl)carbamate(for a preparation see Intermediate 40)(35 mg, 0.064 mmol) in1,4-dioxane (1 mL) at room temperature was treated with hydrogenchloride in 1,4-dioxane (4N, 2 mL, 8.00 mmol) and the resultingsuspension was stirred for 1 h at this temperature then concentrated invacuo. The residue was triturated with Et₂O the solid isolated byfiltration and dried under house vacuum to give4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-aminoethyl)benzamide,hydrochloride (30 mg, 0.062 mmol, 97% yield) as a very pale yellowsolid. LCMS (HpH, 2 min), Rt=0.74 min, MH+=444.

Example 236-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide

To a flask charged with6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)nicotinicacid (for a preparation see Intermediate 28)(50 mg, 0.124 mmol) and HATU(56.7 mg, 0.149 mmol) was added DMF (2 mL) and the resulting mixture wastreated with DIPEA (0.065 mL, 0.373 mmol) then after 5 min withN1,N1-dimethylethane-1,2-diamine (21.90 mg, 0.248 mmol. The resultingsolution was stirred at room temperature for 30 min then partitionedbetween EtOAc and saturated NaHCO₃. The organic phase was separated andaqueous layer extracted with EtOAc (×2). The combined organic layerswere washed with saturated NaHCO₃, dried over MgSO₄ and concentrated invacuo. The residue divided into 2 portions and each was purified by MDAP(HpH). The appropriate fractions were collected and concentrated invacuo. The residue was dissolved in MeOH and concentrated again, thenco-evaporated with toluene. The residue were then dissolved in1,4-dioxane (2 mL) and treated with hydrogen chloride in 1,4-dioxane(4N, 0.5 mL). After 2 min the solvent was evaporated in vacuo and theprecipitate formed triturated with Et₂O then dried under house vacuum togive6-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide,as the d-hydrochloride salt (37 mg, 0.068 mmol, 54.6% yield) as a yellowsolid. LCMS (HpH, 2 min), Rt=0.81 min, MH+=473.

The following analogues were prepared by using the above method with theappropriate amine:—

Example 246-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)nicotinamide

LCMS (HpH, 2 min), Rt=0.73 min, MH+=446.

Example 256-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)nicotinamide

LCMS (HpH, 2 min), Rt=0.69 min, MH+=476.

Example 264-((2S,4R)-1-acetyl-2-methyl-4-(pyrimidin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

To a flask charged with4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide(for a preparation see Intermediate 32) (197 mg, 0.5 mmol),2-bromopyrimidine (238 mg, 1.500 mmol), sodium tert-butoxide (240 mg,2.500 mmol), 2-(dimethylamino)-2′-(dicyclohexylphosphino)biphenyl (197mg, 0.500 mmol) and tris(dibenzylideneacetone)dipalladium(0) (229 mg,0.250 mmol) was added degassed toluene (4 mL) and the resulting mixturewas stirred under nitrogen at 100° C. After 2 h the mixture was cooledto room temperature and most of the solvent was evaporated in vacuo. Theresidue was partitioned between EtOAc and saturated NaHCO₃ solution andthe layers were separated. The aqueous phase was extracted with EtOAcand the combined organic phases were washed with saturated aqueousNaHCO₃ (×3), dried over MgSO₄ and concentrated in vacuo. The residue wasdissolved in DCM and loaded on a SCX column (20 g). The cartridge waseluted with DCM then ammonia in MeOH (2N). The methanolic ammoniafractions were concentrated in vacuo and further purified on bychromatography (25 g column, elution with 2N ammonia in MeOH/DCMgradient) to give a residue which was dissolved in 1,4-dioxane (4 ml)and the solution treated with hydrogen chloride in 1,4-dioxane (4N, 0.5mL). The precipitate which formed was isolated by filtration and driedat 60° C. under vacuum for 2 h to give4-((2S,4R)-1-acetyl-2-methyl-4-(pyrimidin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide,hydrochloride (93 mg, 0.183 mmol, 36.5% yield) as a pale yellow solid.LCMS (HpH, 2 min), Rt=0.79 min, MH+=473.

Example 274-((2S,4R)-1-acetyl-2-methyl-4-(pyrazin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

To a flask charged with4-((2S,4R)-1-acetyl-4-amino-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide(for a preparation see Intermediate 32) (197 mg, 0.5 mmol), sodiumtert-butoxide (240 mg, 2.500 mmol),tris(dibenzylideneacetone)dipalladium(0) (229 mg, 0.250 mmol) and2-(dimethylamino)-2′-(dicyclohexylphosphino)biphenyl (197 mg, 0.500mmol) was added degassed toluene (4 mL). The resulting mixture wastreated with 2-chloropyrazine (0.134 mL, 1.500 mmol) then stirred undernitrogen at 100° C. for ˜45 min. The mixture was cooled to roomtemperature and most of the solvent was evaporated in vacuo. The residuewas partitioned between EtOAc and a saturated NaHCO₃ solution and thelayers were separated. The aqueous phase was extracted with EtOAc andthe combined organic phases were washed with saturated aqueous NaHCO₃(×3), dried over MgSO₄ and concentrated in vacuo. The residue wasdissolved in DCM and loaded on a SCX column (20 g). The cartridge waseluted with DCM then with ammonia in MeOH (2N). The methanolic ammoniafractions were concentrated in vacuo and further purified bychromatography (25 g column, elution with 2N ammonia in MeOH/DCMgradient) to give a residue which was dissolved in 1,4-dioxane (4 ml).The solution was treated with hydrogen chloride in 1,4-dioxane (4N, 0.5mL). The precipitate formed was isolated by filtration and dried at 60°C. under vacuum for 2 h to give4-((2S,4R)-1-acetyl-2-methyl-4-(pyrimidin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide,Hydrochloride (93 mg, 0.183 mmol, 36.5% yield) as a white solid. LCMS(HpH, 2 min), Rt=0.79 min, MH+=473.

Example 285-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)picolinamide

5-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (for a preparation see Intermediate 34) (60 mg, 0.140 mmol) wasdissolved in anhydrous DMF (3 ml) and HATU (64.0 mg, 0.168 mmol) wasadded. DIPEA (0.074 ml, 0.421 mmol) was added and after a further 5 minof stirring at r.t. N1,N1-dimethylethane-1,2-diamine (0.019 ml, 0.281mmol) was added and reaction mixture stirred overnight at r.t. Thereaction mixture was concentrated in vacuo and partitioned between EtOAcand water. The organic phase was separated, dried (Na₂SO₄) andconcentrated to give a yellow residue which was purified bychromatography on a silica cartridge (10 g), eluting with a MeOH/DCMgradient (2-12%, over 120 mls) to give pale yellow solid (28 mg). Thissolid was treated with 1.1 equivalents of 1M hydrogen chloride indiethyl ether and the mixture triturated with diethyl ether to giveyellow solid5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)picolinamide,hydrochloride (32 mg, 0.054 mmol, 38.4% yield).

LC/MS (Formic, 2 min), Rt=0.65 min, MH+=498.

The following analogues were prepared by using the above method with theappropriate amine:—

Example 295-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)picolinamide

LCMS (Formate, 2 min), Rt=0.74 min, MH+=471

Example 305-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)picolinamide

LCMS (Formate, 2 min), Rt=0.70 min, MH+=501

Example 315-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(3-hydroxypropyl)picolinamide

LCMS (Formic, 2 min), Rt=0.77 min, MH+=485

Example 325-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((R)-1-hydroxypropan-2-yl)picolinamide

LCMS (Formate, 2 min), Rt=0.79 min, MH+=485

Example 336-(((2S,4R)-1-acetyl-6-(6-(3-hydroxypyrrolidine-1-carbonyl)pyridin-3-yl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

LCMS (Formate, 2 min), Rt=0.72 min, MH+=497

Example 34 6-(((2S,4R)-1-acetyl-2-methyl-6-(6-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

LCMS (Formate, 2 min), Rt=0.64 min, MH+=510

Example 356-(((2S,4R)-1-acetyl-6-(6-(4-hydroxypiperidine-1-carbonyl)pyridin-3-yl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

LCMS (Formate, 2 min), Rt=0.72 min, MH+=511

Example 366-(((2S,4R)-1-acetyl-6-(6-(4-aminopiperidine-1-carbonyl)pyridin-3-yl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

tert-Butyl(1-(5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinoyl)piperidin-4-yl)carbamate(for a preparation see Intermediate 43) (0.258 ml, 0.313 mmol) wasdissolved in anhydrous DCM (4 ml). TFA (1 ml, 12.98 mmol) was added andthe reaction mixture which was stirred under nitrogen at r.t. for 8 h.The reaction mixture was concentrated in vacuo and loaded onto a SCXcartridge (5 g, pre-conditioned with MeOH). The cartridge was washedwith MeOH (40 ml) and eluted with ammonia in MeOH (2M). The methanolicammonia fractions were concentrated to give an orange oil. The oil wasfurther purified by chromatography on a silica cartridge (10 g), elutingwith a MeOH/DCM gradient (2-12%, over 120 mls) followed by 10% of 2M NH3in MeOH/DCM for 120 mls to give a pale orange solid. The solid wasre-dissolved in MeOH (1 ml) and 1.1 equivalents of 1M hydrogen chloridein diethyl ether were added. The mixture was triturated with diethylether to give6-(((2S,4R)-1-acetyl-6-(6-(4-aminopiperidine-1-carbonyl)pyridin-3-yl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile,hydrochloride (19 mg, 0.030 mmol, 8.59% yield) as an orange solid. LCMS(Formate, 2 min), Rt=0.60 min, MH+=510.

The following analogues were prepared by using the above method:—

Example 376-(((2S,4R)-1-acetyl-6-(6-(3-aminoazetidine-1-carbonyl)pyridin-3-yl)-2-methyl-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

LCMS (Formate, 2 min), Rt=0.60 min, MH+=482

Example 384-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoicacid (for a preparation see Intermediate 18) (150 mg, 0.345 mmol) wassuspended in DCM (3 mL) and DMAP (51.5 mg, 0.422 mmol) then DCC (78.8mg, 0.382 mmol) added. The reaction was stirred for 5 min beforeN,N-dimethyl-ethylenediamine (33.4 mg, 0.379 mmol) in DCM (0.5 mL) wasadded. The reaction was stirred at room temperature overnight. Thereaction was concentrated in vacuo. The residue was taken up in theminimum volume of 5% MeOH in DCM and applied to a 25 g cartridge andallowed to dry in the vac oven at 40° C. for 30 min. The cartridge waseluted with 1% 2M ammonia in Methanol in DCM for 2CV then 1-10% 2Mammonia in Methanol over 10CV then held at 10% for 5CV. The product wasisolated as a yellow solid which was further purified by MDAP (HpH). Theproduct containing fractions were concentrated in vacuo. The residue wastaken up in the minimum volume of MeOH and applied to a SCX cartridge (5g) which was eluted with MeOH (20 mL) then ammonia in MeOH (2M, 20 mL).The ammonia fraction was concentrated in vacuo to yield a clear oil.(85.8 mg, 0.17 mmol). This was taken up in the minimum volume of DCM and1.1 eq 1M HCl in Et₂O (0.19 ml) was added. The solvent was removed undera stream of nitrogen and Et₂O(˜2 ml) added to the residue then removedunder a stream of nitrogen to yield4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}-N42-(dimethylamino)ethypenzamidehydrochloride (77.2 mg, 0.140 mmol, 40.7% yield) as a white solid. LCMS(Formate, 2 min), Rt=0.85 min, MH+=505.

Example 394-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide

A flask was charged with4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}benzoicacid (for a preparation see Intermediate 18) (109 mg, 0.25 mmol), HOBT(45.9 mg, 0.300 mmol) and EDC (57.5 mg, 0.300 mmol) DCM (5 mL) added andthe resulting mixture treated with N-ethylmorpholine (0.095 mL, 0.750mmol). After 5 min, the pale yellow solution was treated with2-amino-1,3-propanediol (29.6 mg, 0.325 mmol) and the mixture stirred atroom temperature. After 4 h further portions of HOBT (22.95 mg, 0.150mmol), EDC (28.75 mg, 0.150 mmol) and N-ethylmorpholine (0.0475 mL,0.375 mmol) were added and reaction mixture stirred overnight at r.t.The volatiles were evaporated in vacuo and the residue dissolved inEtOAc and water. The layers were separated and the aqueous phaseextracted with EtOAc. The combined organic phases were washed withsaturated aqueous NaHCO₃ (×2), then brine, dried over MgSO₄ andconcentrated in vacuo. The residue was purified by chromatographyeluting with a MeOH/DCM gradient to give4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}-N-[2-hydroxy-1-(hydroxymethyl)ethypenzamide(66 mg, 0.130 mmol, 52% yield) as a pale yellow foam. LCMS (HpH, 2 min),Rt=0.95 min, MH+=508.

The following analogues were prepared by using the above method usingthe appropriate amine:—

Example 404-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)benzamide

LCMS (H pH, 2 min), Rt=1.00 min, MH+=478

Example 414-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((R)-2-hydroxypropyl)benzamide

LCMS (High pH, 2 min), Rt=1.03 min, MH+=492

Example 424-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((S)-2-hydroxypropyl)benzamide

LCMS (HpH, 2 min), Rt=1.03 min, MH−=490

Example 434-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N—((S)-2,3-dihydroxypropyl)benzamide

LCMS (HpH, 2 min), Rt=0.95 min, MH+=508

Example 444-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-aminoethyl)benzamide

A solution of 1,1-dimethylethyl(2-{[(4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}phenyl)carbonyl]amino}ethyl)carbamate(for a preparation see Intermediate 45) (57 mg, 0.099 mmol) in1,4-dioxane (0.5 mL) at room temperature was treated with hydrogenchloride in 1,4-dioxane (2 mL, 8.0 mmol) and the resulting mixture wasstirred at room temperature for 1.5 h then concentrated in vacuo. Theresidue was co-evaporated with Et₂O to give4-{(2S,4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1,2,3,4-tetrahydro-6-quinolinyl}-N-(2-aminoethyl)benzamide(32.5 mg, 0.063 mmol, 64.1% yield) hydrochloride as a pale yellow solid.LCMS (HpH, 2 min), Rt=1.01 min, MH+=477.

Example 454-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)benzamide

4-((2S,4R)-1-Acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid (for a preparation see Intermediate 35) (90 mg, 0.225 mmol) wasdissolved in anhydrous DMF (3 ml and HATU (103 mg, 0.270 mmol) added.DIPEA (0.118 ml, 0.674 mmol) was added to the mixture and after afurther 5 min of stirring at r.t. N1,N1-dimethylethane-1,2-diamine(0.030 ml, 0.449 mmol). The reaction mixture was stirred at r.t.overnight, concentrated and partitioned between EtOAc and water. TheOrganic phase was separated, dried (Na₂SO₄) and concentrated. The yellowresidue was purified by chromatography a silica cartridge (10 g),eluting with MeOH/DCM (1-6% over 120 mls, followed by 6-20% over 120mls) to give4-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2(dimethylamino)ethyl)benzamide(24 mg, 0.05 mmol, 20% yield) as a light brown solid. LCMS (Formic, 2min), Rt=0.79 min, MH+=471.

Analogues prepared by the above method using the appropriate amine:—

Examples 464-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)benzamide

LCMS (Formate, 2 min), Rt=0.88 min, MH+=444

Example 474-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide

LCMS (Formate, 2 min), Rt=0.82 min, MH+=474.

Example 486-(((2S,4R)-1-acetyl-2-methyl-6-(6-(piperazine-1-carbonyl)pyridin-3-yl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

tert-Butyl4-(5-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinoyl)piperazine-1-carboxylate(for a preparation see Intermediate 44) (205 mg, 0.344 mmol), wasdissolved in anhydrous DCM (4 ml). TFA (1 ml, 12.98 mmol) was added toeach reaction mixture, which was stirred under nitrogen at r.t. for 7.5h. The reaction mixture was concentrated and loaded onto an SCXcartridge (5 g, pre-conditioned with MeOH). The cartridge was washedwith MeOH (40 ml) followed by ammonia in MeOH (2M). and the methanolicammonia fraction concentrated. The residue was loaded onto a silicacartridge (10 g) and the cartridge washed with 10% MeOH/DCM followed by10% 2M ammonia in MeOH/DCM. The appropriate fractions were concentrated,the residue re-dissolved in MeOH (1 ml) treated with 1.1 equivalents ofhydrogen chloride in diethyl ether (1M). Trituration with ether gave4-((2S,4R)-1-acetyl-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1,3-dihydroxypropan-2-yl)benzamide.hydrochloride as an orange solid (57 mg, 26.5%). LCMS (formate, 2 min)Rt=0.60 mins, 83%, MH+ at 496

Example 495-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)picolinamide

5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)picolinicacid (for a preparation see Intermediate 53) (100 mg, 0.233 mmol), HATU(177 mg, 0.467 mmol) and DIPEA (0.122 ml, 0.700 mmol) were combined inDMF (3 mL) and stirred at room temperature for 10 mins. 2-aminoethanol(0.029 ml, 0.350 mmol) was added and the reaction mixture stirred for1.5 h. The mixture was concentrated in vacuo and purified by MDAP(formate) to give5-((2S,4R)-1-acetyl-4-((5-cyanopyrazin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)picolinamide(42 mg, 0.089 mmol, 38% yield). LCMS: (Formate, 2 min), Rt=0.73 mins,MH+=472

Example 501-((2S,4R)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 56) (40 mg, 0.101 mmol),2-bromopyridine (0.019 mL, 0.203 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (8.0 mg,0.020 mmol), tris(dibenzylideneacetone)dipalladium(0) (9.3 mg, 10.1μmol) and sodium tert-butoxide (19.5 mg, 0.203 mmol) were combined in1,4-dioxane (1 mL), degassed over a period of 15 mins and heated undermicrowave irradiation to 120° C. for 25 mins. A further portion oftris(dibenzylideneacetone)dipalladium(0) (9.3 mg, 10.1 μmol) was added,the reaction degassed, and the mixture heated under microwaveirradiation to 120° C. for 20 mins.

In a separate vial,1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation see Intermediate 56) (25 mg, 0.063 mmol),2-bromopyridine (0.012 mL, 0.127 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (5.0 mg,0.013 mmol), tris(dibenzylideneacetone)dipalladium(0) (5.8 mg, 6.34μmol) and sodium tert-butoxide (12.2 mg, 0.127 mmol) were combined in1,4-dioxane (1 mL), degassed over a period of 15 mins and heated undermicrowave irradiation to 120° C. for 25 mins.

The two crude reaction mixtures were combined and partioned betweensaturated aq. sodium bicarbonate (50 mL) and DCM (3×50 mL). The organicswere combined, dried by passing through a hydrophobic frit andconcentrated in vacuo. The residue was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(36 mg, 0.076 mmol, 75% yield). LCMS: (Formate, 2 min), Rt=0.56 mins,MH+=472.

Example 511-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (80 mg, 0.203 mmol),2-bromo-5-methylpyridine (69.8 mg, 0.406 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (16.0mg, 0.041 mmol), tris(dibenzylideneacetone)dipalladium(0) (18.6 mg,0.020 mmol) and sodium tert-butoxide (39.0 mg, 0.406 mmol) were combinedin 1,4-dioxane (2 mL). The mixture was degassed over a period of 15 minsbefore being heated under microwave irradiation to 120° C. for 30 mins.The mixture was partitioned between saturated aq. sodium bicarbonate (50mL) and DCM (3×50 mL). The organic layers were combined, dried bypassing through a hydrophobic frit and concentrated in vacuo. Theresulting residue was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(13 mg, 0.027 mmol, 13% yield) as an off-white solid. LCMS: (Formate, 2min), Rt=0.58 mins, MH+=486.

Example 521-((2S,4R)-4-((5-chloropyridin-2-yl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (80 mg, 0.203 mmol),2-bromo-5-chloropyridine (78 mg, 0.406 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (16.0mg, 0.041 mmol), tris(dibenzylideneacetone)dipalladium(0) (18.6 mg,0.020 mmol) and sodium tert-butoxide (27.3 mg, 0.284 mmol) were combinedin 1,4-dioxane (1 mL). The mixture was degassed over a period of 15 minsbefore being heated under microwave irradiation to 120° C. for 15 mins.The mixture was partitioned between saturated aq. sodium bicarbonate (50mL) and DCM (3×50 mL). The organic layers were combined, dried bypassing through a hydrophobic frit and concentrated in vacuo. Theresulting residue was purified by MDAP (HpH) to give1-((2S,4R)-4-((5-chloropyridin-2-yl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(54 mg, 0.107 mmol, 53% yield). LCMS: (Formate, 2 min), Rt=0.86 mins,MH+=506.

Example 531-((2S,4R)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-4-(pyrazin-2-ylamino)-3,4-dihydroquinolin-1(2H)yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (80 mg, 0.203 mmol),2-chloropyrazine (0.036 mL, 0.406 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (16.0mg, 0.041 mmol), tris(dibenzylideneacetone)dipalladium(0) (18.6 mg,0.020 mmol) and sodium tert-butoxide (27.3 mg, 0.284 mmol) weredissolved in 1,4-dioxane (2 mL). The mixture was degassed over a periodof 15 mins before being heated under microwave irradiation to 120° C.for 25 mins. The mixture was partitioned between saturated aq. sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organic layers were combined,dried by passing through a hydrophobic frit and concentrated in vacuo.The resulting residue was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-4-(pyrazin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(50 mg, 0.106 mmol, 52% yield).

LCMS: (Formate, 2 min), Rt=0.68 mins, MH+=473.

Example 541-((2S,4R)-2-methyl-4-((5-methylpyrazin-2-yl)amino)-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (80 mg, 0.203 mmol),2-bromo-5-methylpyrazine (70.2 mg, 0.406 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1-biphenyl]-2-amine (16.0 mg,0.041 mmol), tris(dibenzylideneacetone)dipalladium(0) (18.6 mg, 0.020mmol) and sodium tert-butoxide (27.3 mg, 0.284 mmol) were combined in1,4-dioxane (2 mL). The mixture was degassed over a period of 15 minsbefore being heated under microwave irradiation to 120° C. for 25 mins.The mixture was partitioned between saturated aq. sodium bicarbonate (50mL) and DCM (3×50 mL). The organic layers were combined, dried bypassing through a hydrophobic frit and concentrated in vacuo. Theresulting residue was purified by MDAP (HpH) followed by furtherpurification by MDAP (Formate) to give1-((2S,4R)-2-methyl-4-((5-methylpyrazin-2-yl)amino)-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(26 mg, 0.053 mmol, 26% yield).

LCMS: (Formate, 2 min), Rt=0.71 mins, MH+=487.

Example 551-((2S,4R)-2-methyl-6-(4-(morpholine-4-carbonyl)phenyl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate

Crude lithium4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 58) (40 mg) and HATU (41 mg, 0.108mmol) in DMF (0.5 mL) were treated with DIPEA (51.2 μL, 0.294 mmol) andthe mixture shaken and sonicated until a solution was formed (<2 min).This solution was added to morpholine (17 μL, 0.194 mmol), the mixturebriefly shaken then left to stand at room temperature for 2 h. A furtherportion of morpholine (34 μL, 0.388 mmol) was added and the mixture leftto stand overnight. The mixture was diluted with water and extractedthree times with DCM. The combined organic layers were dried by passingthrough a hydrophobic frit and concentrated in vacuo to give a residue.The residue was purified by MDAP (formate) to give1-((2S,4R)-2-methyl-6-(4-(morpholine-4-carbonyl)phenyl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate (20 mg, 0.039 mmol, 10% yield) as a pale pink glass.

LCMS: (Formate, 2 min), Rt 0.63 min, MH+471.

Example 561-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(4-(morpholine-4-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate

A mixture of crude lithium4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 60) (48 mg) and HATU (48 mg, 0.316mmol) in DMF (0.5 mL) was treated with DIPEA (0.06 mL, 0.344 mmol) andthe mixture stirred at room temperature for 5 min to give a solutionwhich was added to morpholine (51 μL, 0.583 mmol). The mixture wasshaken briefly and then left at room temperature overnight. Theresulting solution was diluted with water to and left to stand for ˜10min. The resultant precipitate was isolated by filtration and washedwith water. The isolated precipitate was dissolved in MeOH concentratedto dryness to give a residue which was purified by MDAP (Formate) togive1-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(4-(morpholine-4-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate as a pale yellow glass (24 mg, 0.045 mmol, 8% yield).

LCMS: (Formate, 2 min), Rt=0.66 min, MH+485.

Example 571-((2S,4R)-4-((5-chloropyridin-2-yl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (100 mg, 0.232mmol), 2-bromo-5-chloropyridine (89 mg, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (18.3mg, 0.046 mmol), tris(dibenzylideneacetone)dipalladium(0) (21.3 mg,0.023 mmol) and sodium tert-butoxide (44.6 mg, 0.464 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.A further portion of 2-bromo-5-chloropyridine (89 mg, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (18.3mg, 0.046 mmol), tris(dibenzylideneacetone)dipalladium(0) (21.3 mg,0.023 mmol), sodium tert-butoxide (44.6 mg, 0.464 mmol) and 1,4-dioxane(1 mL) were added and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.The reaction mixture was partitioned between saturated aqueous sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organics were combined, driedby passing through a hydrophobic frit and concentrated in vacuo to givea residue which was purified by MDAP (HpH) to give1-((2S,4R)-4-((5-chloropyridin-2-yl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(34 mg, 0.067 mmol, 29% yield) LCMS: (Formate, 2 min), Rt=0.83 mins,MH+506.

Example 581-((2S,4R)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-4-(pyrazin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (100 mg, 0.232mmol), 2-chloropyrazine (0.041 mL, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (18.3mg, 0.046 mmol), tris(dibenzylideneacetone)dipalladium(0) (21.3 mg,0.023 mmol) and sodium tert-butoxide (44.6 mg, 0.464 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.A further portion of 2-chloropyrazine (0.041 mL, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (18.3mg, 0.046 mmol), tris(dibenzylideneacetone)dipalladium(0) (21.3 mg,0.023 mmol), sodium tert-butoxide (44.6 mg, 0.464 mmol) and 1,4-dioxane(1 mL) were added and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.The reaction mixture was partitioned between saturated aqueous sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organics were combined, driedby passing through a hydrophobic frit and concentrated in vacuo to givea residue which was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-4-(pyrazin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(40 mg, 0.085 mmol, 37% yield). LCMS: (Formate, 2 min), Rt=0.68 mins,MH+473

Example 591-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (80 mg, 0.186mmol), 2-bromo-5-methylpyridine (63.9 mg, 0.371 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (14.6mg, 0.037 mmol), tris(dibenzylideneacetone)dipalladium(0) (17 mg, 0.019mmol) and sodium tert-butoxide (53.5 mg, 0.557 mmol) were combined in1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 130° C. for 50 mins.A further portion of 2-bromo-5-methylpyridine (63.9 mg, 0.371 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (14.6mg, 0.037 mmol), tris(dibenzylideneacetone)dipalladium(0) (17 mg, 0.019mmol), sodium tert-butoxide (53.5 mg, 0.557 mmol) and 1,4-dioxane (1 mL)were added and the mixture degassed over a period of 15 mins, beforebeing heated under microwave irradiation to 130° C. for 25 mins. Thereaction mixture was partitioned between saturated aqueous sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organics were combined, driedby passing through a hydrophobic frit and concentrated in vacuo to givea residue which was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(38 mg, 0.078 mmol, 42% yield). LCMS: (Formate, 2 min), Rt=0.58 mins,MH+486.

Example 601-((2S,4R)-2-methyl-4-((5-methylpyrazin-2-yl)amino)-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (100 mg, 0.232mmol), 2-bromo-5-methylpyrazine (80 mg, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (27.4mg, 0.070 mmol), tris(dibenzylideneacetone)dipalladium(0) (42.5 mg,0.046 mmol) and sodium tert-butoxide (66.9 mg, 0.696 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 130° C. for 40 mins.The reaction mixture was partitioned between saturated aqueous sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organics were combined, driedby passing through a hydrophobic frit and concentrated in vacuo to givea residue which was purified by MDAP (HpH) to give1-((2S,4R)-2-methyl-4-((5-methylpyrazin-2-yl)amino)-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(38 mg, 0.078 mmol, 34% yield). LCMS: (Formate, 2 min), Rt=0.71 mins,MH+487

Example 611-((2S,4R)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (100 mg, 0.232mmol), 2-bromopyridine (0.044 mL, 0.464 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (18.3mg, 0.046 mmol), tris(dibenzylideneacetone)dipalladium(0) (21.3 mg,0.023 mmol) and sodium tert-butoxide (66.9 mg, 0.696 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 130° C. for 25 mins.The reaction mixture was partitioned between saturated aqueous sodiumbicarbonate (50 mL) and DCM (3×50 mL). The organics were combined, driedby passing through a hydrophobic frit and concentrated in vacuo to givea residue which was dissolved in acetonitrile (20 mL) and 0.1M aq.ammonium bicarbonate (20 mL). The mixture was filtered, and the filtrateconcentrated in vacuo to give a residue which was purified by MDAP (HpH)to give1-((2S,4R)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-4-(pyridin-2-ylamino)-3,4-dihydroquinolin-1(2H)-yl)ethanone(41 mg, 0.087 mmol, 38% yield). LCMS: (Formate, 2 min), Rt=0.55 mins,MH+472

Example 621-((2S,4R)-4-(3-chlorophenyl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (100 mg, 0.254 mmol),1-bromo-3-chlorobenzene (0.060 mL, 0.507 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (20.0mg, 0.051 mmol), tris(dibenzylideneacetone)dipalladium(0) (23.3 mg,0.025 mmol) and sodium tert-butoxide (34.1 mg, 0.355 mmol) were combinedin 1,4-dioxane (2.5 mL). The mixture was degassed over a period of 15mins before being heated under microwave irradiation to 120° C. for 25mins. The mixture was partitioned between water (50 mL) and DCM (3×50mL). The organic layers were combined, dried by passing through ahydrophobic frit and concentrated in vacuo. The resulting residue waspurified by MDAP (HpH) to give1-((2S,4R)-4-((3-chlorophenyl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(58 mg, 0.115 mmol, 45% yield).

LCMS: (Formate, 2 min), Rt=1.00 mins, MH+=505

Example 631-((2S,4R)-4-(3-chlorophenyl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (120 mg, 0.278mmol), 1-bromo-3-chlorobenzene (0.065 mL, 0.557 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (21.9mg, 0.056 mmol), tris(dibenzylideneacetone)dipalladium(0) (25.5 mg,0.028 mmol) and sodium tert-butoxide (37.5 mg, 0.390 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.The reaction mixture was partitioned between water (50 mL) and DCM (3×50mL). The organics were combined, dried by passing through a hydrophobicfrit and concentrated in vacuo to give a residue which was purified byMDAP (HpH) to give1-((2S,4R)-4-((3-chlorophenyl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(13 mg, 0.026 mmol, 9.2% yield).

LCMS: (Formate, 2 min), Rt=1.00 mins, MH+505.

Example 641-((2S,4R)-4-((4-chlorophenyl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanonehydrochloride (for a preparation see Intermediate 65) (120 mg, 0.278mmol), 1-bromo-4-chlorobenzene (107 mg, 0.557 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (21.9mg, 0.056 mmol), tris(dibenzylideneacetone)dipalladium(0) (25.5 mg,0.028 mmol) and sodium tert-butoxide (37.5 mg, 0.390 mmol) were combinedin 1,4-dioxane (2 mL) and the mixture degassed over a period of 15 mins,before being heated under microwave irradiation to 120° C. for 25 mins.The reaction mixture was partitioned between water (50 mL) and DCM (3×50mL). The organics were combined, dried by passing through a hydrophobicfrit and concentrated in vacuo to give a residue which was purified byMDAP (HpH) to give1-((2S,4R)-4-((4-chlorophenyl)amino)-2-methyl-6-(5-(morpholine-4-carbonyl)pyridin-2-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(15 mg, 0.030 mmol, 10.7% yield).

LCMS: (Formate, 2 min), Rt=1.00 mins, MH+=505.

Example 656-(((2S,4R)-1-acetyl-2-methyl-6-(4-(morpholine-4-carbonyl)phenyl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile

Crude lithium4-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)benzoicacid. (Intermediate 67) (55 mg) and HATU (53 mg, 0.140 mmol) inanhydrous DMF (1 mL) was treated with DIPEA (67.5 μL, 0.388 mmol). Themixture was sonicated briefly to give a solution, to which was addedmorpholine (110 μL, 1.26 mmol). The solution was left at roomtemperature overnight and then diluted with water to ˜10 mL. The mixturewas further diluted with brine and extracted with DCM/EtOAc and thenEtOAc. The combined organic fractions were dried by passing through ahydrophobic frit and concentrated to dryness under a stream of nitrogen.The resulting residue was purified by MDAP (Formate) to give6-(((2S,4R)-1-acetyl-2-methyl-6-(4-(morpholine-4-carbonyl)phenyl)-1,2,3,4-tetrahydroquinolin-4-yl)amino)nicotinonitrile(26 mg, 0.052 mmol) as a colourless glass. LCMS; (Formate, 2 min)Rt=0.85 min, MH+496.

Example 661-((2S,4R)-4-(5-fluoropyridin-2-yl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone

1-((2S,4R)-4-amino-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(for a preparation, see Intermediate 56) (145 mg, 0.368 mmol),2-bromo-5-fluoropyridine (129 mg, 0.735 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (29 mg,0.074 mmol), tris(dibenzylideneacetone)dipalladium(0) (34 mg, 0.037mmol) and sodium tert-butoxide (71 mg, 0.739 mmol) were combined in1,4-dioxane (4 mL). The mixture was degassed over a period of 15 minsbefore being heated under microwave irradiation to 120° C. for 30 mins.A further portion of 2-bromo-5-fluoropyridine (129 mg, 0.735 mmol),2′-(dicyclohexylphosphino)-N,N-dimethyl-[1,1′-biphenyl]-2-amine (29 mg,0.074 mmol), tris(dibenzylideneacetone)dipalladium(0) (34 mg, 0.037mmol) were added, The mixture was degassed over a period of 15 minsbefore being heated under microwave irradiation to 120° C. for 30 mins.The mixture was partitioned between water (20 mL) and DCM (3×15 mL). Theorganic layers were combined, dried by passing through a hydrophobicfrit and concentrated in vacuo. The resulting residue was purified bychromatography (25 g column, (MeOH/DCM gradient) to give1-((2S,4R)-4-((5-fluoropyridin-2-yl)amino)-2-methyl-6-(6-(morpholine-4-carbonyl)pyridin-3-yl)-3,4-dihydroquinolin-1(2H)-yl)ethanone(91 mg, 0.186 mmol, 51% yield). LCMS: (Formate, 2 min), Rt=0.73 mins,MH+490.

Example 671-((2S,4R)-2-methyl-4-(pyridin-2-ylamino)-6-(4-(pyrrolidine-1-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate

Crude lithium4-((2S,4R)-1-acetyl-2-methyl-4-(pyridin-2-ylamino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoatefor a preparation see Intermediate 58) (40 mg) and HATU (41 mg, 0.108mmol) in DMF (0.5 mL) were treated with DIPEA (51.2 μL, 0.294 mmol) andthe mixture shaken and sonicated until a solution was formed (<2 min).This solution was added to pyrrolidine (16 μL, 0.192 mmol), the mixturebriefly shaken then left to stand at room temperature for 2 h. A furtherportion of morpholine (32 μL, 0.384 mmol) was added and the mixture leftto stand overnight. The mixture was diluted with water and extractedthree times with DCM. The combined organic layers were dried by passingthrough a hydrophobic frit and concentrated in vacuo to give a residue.The residue was purified by MDAP (formate) to give1-((2S,4R)-2-methyl-4-(pyridin-2-ylamino)-6-(4-(pyrrolidine-1-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate (23 mg, 0.046 mmol, 12%) as a pale pink glass. LCMS: (Formate, 2min), Rt=0.70 min, MH+455

Example 681-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(4-(pyrrolidine-1-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate

A mixture of crude lithium4-((2S,4R)-1-acetyl-2-methyl-4-((5-methylpyridin-2-yl)amino)-1,2,3,4-tetrahydroquinolin-6-yl)benzoate(for a preparation see Intermediate 60) (48 mg) and HATU (48 mg, 0.316mmol) in DMF (0.5 mL) was treated with DIPEA (0.06 mL, 0.344 mmol) andthe mixture stirred at room temperature for 5 min to give a solutionwhich was added to pyrollidine (48 μL, 0.575 mmol). The mixture wasshaken briefly and then left at room temperature overnight. Theresulting solution was diluted with water to and left to stand for ˜10min. The resultant precipitate was isolated by filtration and washedwith water. The isolated precipitate was dissolved in MeOH concentratedto dryness to give a residue which was purified by MDAP (Formate) togive1-((2S,4R)-2-methyl-4-((5-methylpyridin-2-yl)amino)-6-(4-(pyrrolidine-1-carbonyl)phenyl)-3,4-dihydroquinolin-1(2H)-yl)ethanoneformate (22 mg, 0.047 mmol, 7.5%) as a pale yellow glass.

LCMS: (Formate, 2 min), Rt=0.73 min, MH+469.

REFERENCE COMPOUNDS

Experimental details of LC-MS methods A-F as referred to herein are asfollows:

LC/MS (Method A) was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm i.d. 1.7 μm packing diameter) at 40 degrees centigrade,eluting with 10 mM Ammonium Bicarbonate in water adjusted to pH 10 withAmmonia solution (Solvent A) and Acetonitrile (Solvent B) using thefollowing elution gradient 0-1.5 min 1-97% B, 1.5-1.9 min 97% B, 1.9-2.0min 100% B at a flow rate of 1 ml/min. The UV detection was a summedsignal from wavelength of 210 nm to 350 nm. The mass spectra wererecorded on a Waters ZQ Mass Spectrometer using Alternate-scan Positiveand Negative Electrospray. Ionisation data was rounded to the nearestinteger.

LC/MS (Method B) was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm i.d. 1.7 μm packing diameter) at 40 degrees centigrade,eluting with 0.1% v/v solution of Formic Acid in Water (Solvent A) and0.1% v/v solution of Formic Acid in Acetonitrile (Solvent B) using thefollowing elution gradient 0-1.5 min 3-100% B, 1.5-1.9 min 100% B,1.9-2.0 min 3% B at a flow rate of 1 ml/min. The UV detection was asummed signal from wavelength of 210 nm to 350 nm. The mass spectra wererecorded on a Waters ZQ Mass Spectrometer using Alternate-scan Positiveand Negative Electrospray. Ionisation data was rounded to the nearestinteger.

LC/MS (Method C) was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm i.d. 1.7 μm packing diameter) at 40 degrees centigrade,eluting with 0.1% v/v solution of Trifluoroacetic Acid in Water (SolventA) and 0.1% v/v solution of Trifluoroacetic Acid in Acetonitrile(Solvent B) using the following elution gradient 0-1.5 min 3-100% B,1.5-1.9 min 100% B, 1.9-2.0 min 3% B at a flow rate of 1 ml/min. The UVdetection was a summed signal from wavelength of 210 nm to 350 nm. Themass spectra were recorded on a Waters ZQ Mass Spectrometer usingPositive Electrospray. Ionisation data was rounded to the nearestinteger.

LC/MS (Method D) was conducted on a Supelcosil LCABZ+PLUS column (3 μm,3.3 cm×4.6 mm ID) eluting with 0.1% HCO₂H and 0.01 M ammonium acetate inwater (solvent A), and 95% acetonitrile and 0.05% HCO₂H in water(solvent B), using the following elution gradient 0-0.7 minutes 0% B,0.7-4.2 minutes 0→100% B, 4.2-5.3 minutes 100% B, 5.3-5.5 minutes 100→0%B at a flow rate of 3 mL/minute. The mass spectra (MS) were recorded ona Fisons VG Platform mass spectrometer using electrospray positiveionisation [(ES+ve to give [M+H]⁺ and [M+NH₄]⁺ molecular ions] orelectrospray negative ionisation [(ES-ve to give [M−H]− molecular ion]modes. Analytical data from this apparatus are given with the followingformat: [M+H]⁺ or [M−H]⁻.

LC/MS (Method E) was conducted on a Chromolith Performance RP 18 column(100×4.6 mm id) eluting with 0.01M ammonium acetate in water (solvent A)and 100% acetonitrile (solvent B), using the following elution gradient0-4 minutes 0-100% B, 4-5 minutes 100% B at a flow rate of 5 ml/minute.The mass spectra (MS) were recorded on a micromass Platform-LC massspectrometer using atmospheric pressure chemical positive ionisation[AP+ve to give MH+ molecular ions] or atmospheric pressure chemicalnegative ionisation [AP−ve to give (M−H)− molecular ions] modes.Analytical data from this apparatus are given with the following format:[M+H]+ or [M−H]−.

LC/MS (Method F) was conducted on an Sunfire C18 column (30 mm×4.6 mmi.d. 3.5 μm packing diameter) at 30 degrees centigrade, eluting with0.1% v/v solution of Trifluoroacetic Acid in Water (Solvent A) and 0.1%v/v solution of Trifluoroacetic Acid in Acetonitrile (Solvent B) usingthe following elution gradient 0-0.1 min 3% B, 0.1-4.2 min 3-100% B,4.2-4.8 min 100% B, 4.8-4.9 min 100-3% B, 4.9-5.0 min 3% B at a flowrate of 3 ml/min. The UV detection was an averaged signal fromwavelength of 210 nm to 350 nm and mass spectra were recorded on a massspectrometer using positive electrospray ionization. Ionisation data wasrounded to the nearest integer.

LC/MS (Method G) was conducted on an Acquity UPLC BEH C18 column (50mm×2.1 mm i.d. 1.7 μm packing diameter) at 40 degrees centigrade,eluting with 0.1% v/v solution of Formic Acid in Water (Solvent A) and0.1% v/v solution of Formic Acid in Acetonitrile (Solvent B) using thefollowing elution gradient 0-1.5 min 1-97% B, 1.5-1.9 min 97% B, 1.9-2.0min 97 to 100% B at a flow rate of 1 ml/min. The UV detection was asummed signal from wavelength of 210 nm to 350 nm. The mass spectra wererecorded on a Waters ZQ Mass Spectrometer using Alternate-scan Positiveand Negative Electrospray. Ionisation data was rounded to the nearestinteger.

LC/HRMS: Analytical HPLC was conducted on a Uptisphere-hsc column (3 μm33×3 mm id) eluting with 0.01M ammonium acetate in water (solvent A) and100% acetonitrile (solvent B), using the following elution gradient0-0.5 minutes 5% B, 0.5-3.75 minutes 5→100% B, 3.75-4.5 100% B, 4.5-5100→5% B, 5-5.5 5% B at a flow rate of 1.3 mL/minute. The mass spectra(MS) were recorded on a micromass LCT mass spectrometer usingelectrospray positive ionisation [ES+ve to give MH⁺ molecular ions] orelectrospray negative ionisation [ES−ve to give (M−H)− molecular ions]modes.

TLC (thin layer chromatography) refers to the use of TLC plates sold byMerck coated with silica gel 60 F254.

Reference Compound A 2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one

A solution of 5-methoxyanthranilic acid (Lancaster) (41.8 g, 0.25 mol)was refluxed in acetic anhydride (230 mL) for 3.5 h before beingconcentrated under reduced pressure. The crude compound was thenconcentrated twice in the presence of toluene before being filtered andwashed twice with ether to yield to the title compound (33.7 g, 71%yield) as a brown solid. LC/MS (Method D): m/z 192 [M+H]⁺, Rt 1.69 min.

Reference Compound B[2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone

To a solution of 2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one (for apreparation see Reference compound A) (40.0 g, 0.21 mol) in atoluene/ether (2/1) mixture (760 mL) at 0° C. was added dropwise asolution of 4-chlorophenylmagnesium bromide (170 mL, 1M in Et₂O, 0.17mol). The reaction mixture was allowed to warm to room temperature andstirred for 1 h before being quenched with 1N HCl (200 mL). The aqueouslayer was extracted with EtOAc (3×150 mL) and the combined organics werewashed with brine (100 mL), dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The crude compound was then dissolved in EtOH(400 mL) and 6N HCl (160 mL) was added. The reaction mixture wasrefluxed for 2 h before being concentrated to one-third in volume. Theresulting solid was filtered and washed twice with ether before beingsuspended in EtOAc and neutralised with 1N NaOH. The aqueous layer wasextracted with EtOAc (3×150 mL) and the combined organics were washedwith brine (150 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure. The title compound was obtained as a yellow solid (39g, 88% yield);

LC/MS (Method D): m/z 262 [M+H]+, Rt 2.57 min.

Reference Compound C MethylN¹-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N²-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-α-asparaginate

Methyl N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-α-aspartyl chloride(Int. J. Peptide Protein Res. 1992, 40, 13-18) (93 g, 0.24 mol) wasdissolved in CHCl₃ (270 mL) and[2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone (for apreparation see Reference compound B) (53 g, 0.2 mol) was added. Theresulting mixture was stirred at 60° C. for 1 h before being cooled andconcentrated to 60% in volume. Ether was added at 0° C. and theresulting precipitate was filtered and discarded. The filtrate wasconcentrated under reduced pressure and used without furtherpurification.

Reference Compound D Methyl[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]acetate

To a solution of methylN1-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N2-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-α-asparaginate(for a preparation see Reference compound C) (assumed 0.2 mol) in DCM(500 mL) was added Et₃N (500 mL, 3.65 mol) and the resulting mixture wasrefluxed for 24 h before being concentrated. The resulting crude aminewas dissolved in 1,2-DCE (1.5 L) and AcOH (104 mL, 1.8 mol) was addedcarefully. The reaction mixture was then stirred at 60° C. for 2 hbefore being concentrated in vacuo and dissolved in DCM. The organiclayer was washed with 1N HCl and the aqueous layer was extracted withDCM (×3). The combined organic layers were washed twice with water, andbrine, dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude solid was recrystallised in MeCN leading to thetitle compound (51 g) as a pale yellow solid. The filtrate could beconcentrated and recrystallised in MeCN to give to another 10 g of thedesired product R_(f)=0.34 (DCM/MeOH: 95/5).

HRMS (M+H)⁺ calculated for C₁₉H₁₈ ³⁵ClN₂O₄ 373.0955. found 373.0957.

Reference Compound E Methyl[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]acetate

A suspension of P₄S₁₀ (36.1 g, 81.1 mmol) and Na₂CO₃ (8.6 g, 81.1 mmol)in 1,2-DCE (700 mL) at room temperature was stirred for 2 h beforeMethyl[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]acetate(for a preparation see Reference compound D) (16.8 g, 45.1 mmol) wasadded. The resulting mixture was stirred at 70° C. for 2 h before beingcooled and filtered. The solid was washed twice with DCM and thefiltrate washed with sat. NaHCO₃ and brine. The organic layer was driedover Na₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by flash-chromatography on silica gel (DOM/MeOH:99/1) to afford the title compound (17.2 g, 98% yield) as a yellowishsolid. LC/MS (Method D): m/z 389 [M(³⁵Cl)+H]⁺, Rt 2.64 min

HRMS (M+H)⁺ calculated for C₁₉H₁₈ ³⁵ClN₂O₃S 389.0727. found 389.0714.

Reference Compound F Methyl[(3S)-2-[(1Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H-1,4-benzodiazepin-3-yl]acetate

To a suspension of methyl[(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]acetate(for a preparation see Reference compound E (9.0 g, 23.2 mmol) in THF(300 mL) at 0° C. was added hydrazine monohydrate (3.4 mL, 69.6 mmol)dropwise. The reaction mixture was stirred for 5 h between 5° C. and 15°C. before being cooled at 0° C. Et₃N (9.7 mL, 69.6 mmol) was then addedslowly and acetyl chloride (7.95 mL, 69.6 mmol) was added dropwise. Themixture was then allowed to warm to room temperature for 16 h beforebeing concentrated under reduced pressure. The crude product wasdissolved in DCM and washed with water. The organic layer was dried overNa₂SO₄, filtered and concentrated in vacuo to give the crude titlecompound (9.7 g, 98% yield) which was used without further purification.R_(f)=0.49 (DCM/MeOH:90/10).

Reference Compound G Methyl[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate

The crude methyl[(3S)-2-[(1Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H-1,4-benzodiazepin-3-yl]acetate(for a preparation see Reference compound F) (assumed 9.7 g) wassuspended in THF (100 ml) and AcOH (60 mL) was added at roomtemperature. The reaction mixture was stirred at this temperature for 2days before being concentrated under reduced pressure. The crude solidwas triturated in i-Pr₂O and filtered to give the title compound (8.7 g,91% over 3 steps) as an off-white solid.

HRMS (M+H)⁺ calculated for O₂₁H₂₀ClN₄O₃ 411.1229. found 411.1245.

Reference Compound H[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]aceticacid

To a solution of methyl[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate(for a preparation see Reference compound G)(7.4 g, 18.1 mmol) in THF(130 mL) at room temperature was added 1N NaOH (36.2 mL, 36.2 mmol). Thereaction mixture was stirred at this temperature for 5 h before beingquenched with 1N HCl (36.2 mL) and concentrated in vacuo. Water is thenadded and the aqueous layer was extracted with DCM (×3) and the combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the title compound (7 g, 98% yield) as a paleyellow solid.

LC/MS (Method D): m/z 397 [M+H]⁺

Reference Compound I 1,1-dimethylethyl[5-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetyl}amino)pentyl]carbamate

A mixture of[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]aceticacid (for a preparation see Reference compound H) (1.0 g, 2.5 mmol),HATU (1.9 g, 5 mmol) and DIPEA (0.88 ml, 5 mmol) was stirred for 80minutes at room temperature, to this was added 1,1-dimethylethyl(4-aminobutyl)carbamate (1.05 ml, 5.0 mmol, available from Aldrich). Thereaction mixture was stirred at room temperature for 2 h before it wasconcentrated. The residue was taken up in dichloromethane and washedwith 1N HCl. The aqueous layer was extracted with dichloromethane twice.Organic layer was washed with 1N sodium hydroxide, followed by asaturated solution of sodium chloride, dried over sodium sulphate andconcentrated. The residue was purified by flash-chromatography on silicausing dichloromethane/methanol 95/5 to give the title compound as ayellow solid (1.2 g). LC/MS (Method D): rt=3.04 min.

Reference Compound JN-(5-aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamidetrifluoroacetate

To a solution of 1,1-dimethylethyl[5-({[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetyl}amino)pentyl]carbamate(for a preparation see Reference compound I) (0.2 g, 0.34 mmol) indichloromethane (3 ml) was added trifluoroacetic acid (0.053 ml, 0.68mmol) dropwise at 0° C. The reaction mixture was stirred for 3 h from 0°C. to room temperature. The reaction mixture was concentrated to drynessto afford the title compound as a hygroscopic yellow oil (200 mg)

LC/MS (Method D): rt=2.33 min.

HRMS (M+H)⁺ calculated for C₂₅H₂₉ClN₆O₂ 481.2119. found 481.2162.

Reference Compound K Mixture of 5- and 6-isomers of Alexa Fluor488-N-(5-aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamide

N-(5-Aminopentyl)-2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetamidetrifluoroacetate (for a preparation see Reference compound J) (7.65 mg,0.013 mmol) was dissolved in N,N-Dimethylformamide (DMF) (300 μl) andadded to Alexa Fluor 488 carboxylic acid succinimidyl ester (5 mg, 7.77μmol, mixture of 5 and 6 isomers, available from Invitrogen, productnumber A-20100) in an Eppendorf centrifuge tube. Hunig's base (7.0 μl,0.040 mmol) was added and the mixture vortex mixed overnight. After 18 hthe reaction mixture was evaporated to dryness and the residueredissolved in DMSO/water (50%, <1 ml total), applied to a preparativePhenomenex Jupiter C18 column and eluted with a gradient of 95% A: 5% Bto 100% B (A=0.1% trifluoroacetic acid in water, B=0.1% TFA/90%acetonitrile/10% water) at a flow rate of 10 ml/min over 150 minutes.Impure fractions were combined and re-purified using the same system.Fractions were combined and evaporated to yield the title product (2.8mg) as a mixture of the 2 regioisomers shown. LC/MS (Method F):,MH+=999, rt=1.88 min.

Biological Test Methods Fluorescence Anisotropy Binding Assay

The binding of the compounds of formula (I) to Bromodomains BRD2, BRD3and BRD4 may be assessed using a Fluorescence Anisotropy Binding Assay.

The Bromodomain protein, fluorescent ligand (Reference compound K seeabove) and a variable concentration of test compound are incubatedtogether to reach thermodynamic equilibrium under conditions such thatin the absence of test compound the fluorescent ligand is significantly(>50%) bound and in the presence of a sufficient concentration of apotent inhibitor the anisotropy of the unbound fluorescent ligand ismeasurably different from the bound value.

All data was normalized to the mean of 16 high and 16 low control wellson each plate. A four parameter curve fit of the following form was thenapplied:y=a+((b−a)/(1+(10^x/10^c)^d)

Where ‘a’ is the minimum, ‘b’ is the Hill slope, ‘c’ is the pIC50 and‘d’ is the maximum.

Recombinant Human Bromodomains (BRD2 (1-473), BRD3 (1-435) and BRD4(1-477)) were expressed in E. coli cells (in pET15b vector) with asix-His tag at the N-terminal. The His-tagged Bromodomain was extractedfrom E. coli cells using 0.1 mg/ml lysozyme and sonication. TheBromodomain was then purified by affinity chromatography on a HisTRAP HPcolumn, eluting with a linear 10-500 mM Imidazole gradient, over 20 Cv.Further purification was completed by Superdex 200 prep grade sizeexclusion column. Purified protein was stored at −80 C in 20 mM HEPES pH7.5 and 100 mM NaCl.

Protocol for Bromodomain BRD2:

All components were dissolved in buffer composition of 50 mM HEPESpH7.4, 150 mm NaCl and 0.5 mM CHAPS with final concentrations of BRD2,75 nM, fluorescent ligand 5 nM. 10 μl of this reaction mixture was addedusing a micro multidrop to wells containing 100 nl of variousconcentrations of test compound or DMSO vehicle (1% final) in Greiner384 well Black low volume microtitre plate and equilibrated in dark 60mins at room temperature. Fluorescence anisotropy was read in Envision(λex=485 nm, λEM=530 nm; Dichroic −505 nM).

Protocol for Bromodomain BRD3:

All components were dissolved in buffer of composition 50 mM HEPESpH7.4, 150 mm NaCl and 0.5 mM CHAPS with final concentrations of BRD3 75nM, fluorescent ligand 5 nM. 10 μl of this reaction mixture was addedusing a micro multidrop to wells containing 100 nl of variousconcentrations of test compound or DMSO vehicle (1% final) in Greiner384 well Black low volume microtitre plate and equilibrated in dark 60mins at room temperature. Fluorescence anisotropy was read in Envision(λex=485 nm, λEM=530 nm; Dichroic −505 nM).

Protocol for Bromodomain BRD4:

All components were dissolved in buffer of composition 50 mM HEPESpH7.4, 150 mm NaCl and 0.5 mM CHAPS with final concentrations of BRD4 75nM, fluorescent ligand 5 nM. 10 ml of this reaction mixture was addedusing a micro multidrop to wells containing 100 nl of variousconcentrations of test compound or DMSO vehicle (1% final) in Greiner384 well Black low volume microtitre plate and equilibrated in dark 60mins at room temperature. Fluorescence anisotropy was read in Envision(λex=485 nm, λEM=530 nm; Dichroic −505 nM).

Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) Assay

The binding of the compounds of formula (I) to Bromodomains BRD2, BRD3and BRD4 was assessed using a time resolved fluorescent resonance energytransfer binding assay, that measures the binding of an acetylatedhistone peptide to the bromodomain protein.

The bromodomain protein, histone peptide and a variable concentration oftest compound are incubated together to reach thermodynamic equilibrium.The assay is configured such that in the absence of test compound thebromodomain and peptide are significantly bound (˜30%) and in thepresence of a sufficient concentration of a potent inhibitor thisinteraction is disrupted leading to a measurable drop in fluorescentresonance energy transfer.

Histone Peptide:

H-Ser-Gly-Arg-Gly-Lys(Ac)-Gly-Gly-Lys(Ac)-Gly-Leu-Gly-Lys(Ac)-Gly-Gly-Ala-Lys(Ac)-Arg-His-Gly-Ser-Gly-Ser-Lys(Biotin)-OH. 3TFA

The protected peptide was assembled on a solid-phase synthesiser usingpreloaded Wang resin and utilising standard Fmoc synthesis protocols.The C-terminal lysine was protected by a hyper acid-labile groupallowing for its selective removal at the end of the assembly andattachment of the biotin. The crude peptide was obtained after cleavagefrom the resin with a mixture of trifluoroacetic acid (TFA),triisopropylsilane and water (95:2.5:2.5) for 3 h at room temperatureand was then purified using a C18 reverse-phase column utilising a 0.1%TFA-buffered water/acetonitrile gradient. The resulting fractions wereanalysed and fractions which were >95% pure by analytical HPLC andgiving the correct mw (by MALDiTOF mass spectroscopy) were pooled andfreeze dried. The final material was analysed by HPLC to confirm purity.

Protein Production:

Recombinant Human Bromodomains (BRD2 (1-473), BRD3 (1-435) and BRD4(1-477)) were expressed in E. coli cells (in pET15b vector) with asix-His tag at the N-terminal. The His-tagged Bromodomain was extractedfrom E. coli cells using sonication and purified using a nickelsepharose 6FF column, the proteins were washed and then eluted with 50mM Tris-Hcl pH8.0. 300 mM NaCl, 1 mM 3-mercaptoethanol and 20 mMImidazole. Further purification was performed by affinity chromatographyon a HisTRAP HP column, eluting with a linear 0-500 mM sodium chloridegradient, over 20 column volumes. Final purification was completed bySuperdex 200 prep grade size exclusion column. Purified protein wasstored at −80 C in 20 mM HEPES pH 7.5 and 100 mM NaCl. Protein identitywas confirmed by peptide mass fingerprinting and predicted molecularweight confirmed by mass spectrometry.

Protocol for Bromodomain BRD2, 3 and 4 Assays:

All assay components were dissolved in buffer composition of 50 mM HEPESpH7.4, 50 mM NaCl and 0.5 mM CHAPS. The final concentration ofbromodomain proteins were 100 nM and the histone peptide was 300 nM,these components are premixed and allowed to equilibrate for 1 hour inthe dark. 8 μl of this reaction mixture was added to all wellscontaining 50 nl of various concentrations of test compound or DMSOvehicle (0.5% final) in Greiner 384 well black low volume microtitreplates and incubated in dark for 60 mins at room temperature. 2 μl ofdetection mixture containing anti-6his XL665 labeled antibody andstreptavidin labeled with europium cryptate was added to all wells and afurther dark incubation of at least 30 mins was performed. Plates werethen read on the Envision platereader, (λex=317 nm, donor λEM=615 nm;acceptor λEM=665 nm; Dichroic LANCE dual). Time resolved fluorescentintensity measurements were made at both emission wavelengths and theratio of acceptor/donor was calculated and used for data analysis. Alldata was normalized to the mean of 16 high and 16 low control wells oneach plate. A four parameter curve fit of the following form was thenapplied:y=a+((b−a)/(1+(10^x/10^c)^d)

Where ‘a’ is the minimum, ‘b’ is the Hill slope, ‘c’ is the pIC50 and‘d’ is the maximum.

Examples 1-68 were tested in one or more of the above assays and werefound to have a pIC50 in the range 6.0 to 7.5. Examples 1-5, 7, 10-19,26, 28-31, 39-47, 49, 52 and 65 were found to have a pIC50 in the range7.0-7.5 in at least one of the above assays.

Measurement of LPS Induced IL-6 Secretion from Whole Blood

Activation of monocytic cells by agonists of toll-like receptors such asbacterial lipopolysaccharide (LPS) results in production of keyinflammatory mediators including IL-6. Such pathways are widelyconsidered to be central to the pathophysiology of a range ofauto-immune and inflammatory disorders.

Compounds to be tested are diluted to give a range of appropriateconcentrations of which 1 μl of the diluted stocks is added to a 96 wellplate. Following addition of whole blood (130 μl) the plates areincubated at 37 degrees (5% CO2) for 30 min before the addition of 10 μlof 2.8 ug/ml LPS, diluted in complete RPMI 1640 (final concentration=200ng/ml), to give a total volume of 140 μl per well. After furtherincubation for 24 hours at 37 degrees, 140 μl of PBS are added to eachwell. The plates are sealed, shaken for 10 minutes and then centrifuged(2500 rpm×10 min). 100 μl of the supernatant are removed and IL-6 levelsassayed by immunoassay (typically by MesoScale Discovery technology)either immediately or following storage at −20 degrees. Concentrationresponse curves for each compound was generated from the data and anIC₅₀ value was calculated.

Examples 2, 3, 5-7, 9-12, 14-16, 19-25, 28-32, 34, 37, 38, 40, 42,44-46, 48-49, 51-52, 54, 56-65, and 68 were tested in the above assayand were found to have a pIC50 in the range 5.5 to 7.3.

These data demonstrate that bromodomain inhibitors tested in the abovewhole blood assay inhibited the production of key inflammatory mediatorIL-6.

In Vivo Mouse Endotoxemia Model

High doses of Endotoxin (bacterial lipopolysaccharide) administered toanimals produce a profound shock syndrome including a stronginflammatory response, dysregulation of cardiovascular function, organfailure and ultimately mortality. This pattern of response is verysimilar to human sepsis and septic shock, where the body's response to asignificant bacterial infection can be similarly life threatening.

To test the compounds for use in the invention groups of eight Balb/cmale mice are given a lethal dose of 15 mg/kg LPS by intraperitonealinjection. Ninety minutes later, animals were dosed intravenously withvehicle (20% cyclodextrin 1% ethanol in apyrogen water) or compound (10mg/kg). The survival of animals is monitored at 4 days.

Oncology Cell Growth Assay

Human cell lines (n=33 comprising 15 heme cell lines, 14 breast celllines and 4 other cell lines) were cultured in RPMI-1640 containing 10%fetal bovine serum, 1000 viable cells per well were plated in 384-wellblack flat bottom polystyrene plates (Greiner #781086) in 48 μl ofculture media. All plates were placed at 5% CO₂, 37° C. overnight. Thefollowing day one plate was harvested with CellTiter-Glo (CTG, Promega#G7573) for a time equal to 0 (T0) measurement and compound (20 pointtitration from 14.7 uM to 7 μM) was added to the remaining plates. Thefinal concentration of DMSO in all wells was 0.15%. Cells were incubatedfor 72 hours or the indicated time and each plate was developed withCellTiter-Glo reagent using a volume equivalent to the cell culturevolume in the wells. Plates were shaken for approximately 2 minutes andchemiluminescent signal was read on the Analyst GT (Molecular Devices)or EnVision Plate Reader (Perkin Elmer).

Results are expressed as a percent of the T0 and plotted against thecompound concentration. The T0 value was normalized to 100% andrepresents the number of cells at time of compound addition and theconcentration response data were fit with a 4 parameter curve fit usingXLfit software (model 205). The concentration that inhibited cell growthby 50% (gIC₅₀) is the midpoint of the ‘growth window’ (between the T0and DMSO control). The Ymin−T0 value is determined by subtracting the T0value (100%) from the Ymin value (%) determined from the fit of theconcentration response curve. Values from the wells with no cells weresubtracted from all samples for background correction.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

What is claimed is:
 1. A compound of formula (I) or a salt thereof

in which X and Y are independently CH or N provided that at least one ofX and Y must be CH; R¹ is a group C(O)OR⁴ in which R⁴ is C₁₋₃alkyl orC₃₋₇cycloalkyl; or R¹ is a group selected from phenyl, pyridyl,pyrazinyl and pyrimidinyl said groups being optionally substituted byone or two substituents selected from halogen, C₁₋₄alkyl and CN; R² isC₁₋₄alkyl; R³ is C₁₋₄alkyl; R⁵ is hydrogen and R⁶ is C₁₋₄alkylsubstituted by one or more hydroxy or a —NR⁷R⁸ group in which R⁷ and R⁸are independently hydrogen or a C₁₋₄alkyl group; or R⁵ and R⁶ togetherwith the N to which they are attached form a saturated 4, 5 or 6membered heterocyclyl ring optionally containing a further heteroatomselected from N, O and S, said heterocyclyl ring being optionallysubstituted by one or more Cl_(—)4 alkyl, hydroxyl or amino groups; andm is 0, 1 or
 2. 2. A compound or a salt thereof according to claim 1 inwhich X and Y are both CH.
 3. A compound or a salt thereof according toclaim 1 in which X is CH and Y is N.
 4. A compound or a salt thereofaccording to claim 1 in which R¹ is a group C(O)OR⁴ wherein R⁴ isisopropyl.
 5. A compound of salt thereof according to claim 1 in whichR¹ is selected from


6. A compound or a salt thereof according to claim 1 in which R² ismethyl.
 7. A compound or a salt thereof according to claim 1 in which R³is methyl.
 8. A compound or salt thereof according to claim 1 in which mis
 0. 9. A compound or salt thereof according to claim 1 in which mis
 1. 10. A compound or salt thereof according to claim 1 in which R⁵ ishydrogen and R⁶ is selected from


11. A compound or a salt thereof according to claim 1 in which R⁵ and R⁶combine together with the N to which they are attached to form a groupselected from


12. A compound or a salt thereof according to claim 1 wherein thecompound of formula (I) is the (2S,4R) enantiomer.
 13. A compound whichis selected from2-(4-((2S,4R)-1-Acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-N-(2-(dimethylamino)ethyl)acetamide;6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-(dimethylamino)ethyl)nicotinamide;and6-((2S,4R)-1-acetyl-4-((5-cyanopyridin-2-yl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(2-hydroxyethyl)nicotinamideor a salt thereof.
 14. A compound according to claim 1 or apharmaceutically acceptable salt thereof.
 15. A pharmaceuticalcomposition which comprises a compound or a pharmaceutically acceptablesalt thereof as defined in claim 14 and one or more pharmaceuticallyacceptable carriers, diluents or excipients.
 16. A method of treatingdiseases or conditions for which a bromodomain inhibitor is indicated ina subject in need thereof which comprises administering atherapeutically effective amount of a compound or a pharmaceuticallyacceptable salt thereof as defined in claim 14, wherein the disease orcondition is cancer.